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FAQ
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature/ primer concentration. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (3 ° to 5 °C lower than Tm). Optimise annealing temperature using temperature gradient PCR. Increase primer concentration for lower yield amplicons.
b) Inadequate cycling conditions. Increase extension time and/or number of cycles.
c) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
d) Inadequate amount of template DNA. Titrate template amount. We recommend starting with 20-50 ng of genomic DNA.
c) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.25 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another pool of primers, avoiding complementary sequences. Ensure that each primer pair originates a single product without primer-dimers in individual PCR reactions. If necessary, reduce primer concentration. Reduce the number of cycles.
What is the maximum size of amplicon using Multiplex PCR NZYTaq 2x Colourless Master Mix?
Multiplex PCR NZYTaq 2x Colourless Master Mix allows efficient multiplexing of targets ranging in size from 70 bp to 2.5 k.
Which type of DNA ends are generated in the PCR products using Multiplex PCR NZYTaq 2x Colourless Master Mix?
Resulting PCR products have an A-overhang and are suitable for cloning with NZYTech´s NZY-A PCR cloning kit (MB053) or NZY-A Speedy PCR cloning kit (MB137).
I don’t distinguish separate bands using agarose gel electrophoresis. What should I do?
When separating samples through agarose gel electrophoresis, please take into account that both concentration and grade of agarose are important when working with closely-sized fragments. NZYTech offers a vast portfolio of agaroses with different specifications to cover a wide range of needs. Please visit www.nzytech.com/products-services/category/molecular-biology/products/agaroses/ to identify the agarose that most suits your experiment.
How should I store my Multiplex PCR NZYTaq 2x Colourless Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What information do I need to provide to request a peptide synthesis quotation?
You should provide the peptide sequence, desired quantity, purity level, and any modifications. Please complete the Order Form and submit it via your NZYtech account or directly by email to services@nzytech.com.
What peptide purities are available and how should I choose the right one?
We offer peptides in crude, desalted, and purified formats (>75%, >85%, >95%, >98%). The choice depends on the intended application:
- Crude/desalted: high-throughput or preliminary screening
- >75%: affinity purification, enzyme substrate studies, epitope mapping
- >85%: immunological applications, polyclonal antibody production
- >95%: in vitro bioassays such as ELISA, enzymology
- >98%: structural studies (e.g., NMR, crystallography)
What quantities of peptides can I order?
Standard quantities range from 1-3 mg to 1000 mg. Larger amounts are available upon request.
Which peptide modifications can be introduced?
We provide a wide range of modifications, including phosphorylation, methylation, acetylation, amidation, and fluorescent dye labeling. A complete list of modifications is available in the peptide synthesis modification list available in the product manuals section.
Is it possible to request a salt exchange for my peptide?
Yes, some applications, such as in vivo studies may benefit from TFA-free peptides. Upon request, we can perform a salt exchange to replace TFA (trifluoroacetate) with either HCl (hydrochloride) or acetate salts for an additional fee.
How are the peptides delivered?
All peptides are supplied in lyophilized format.
What is the typical turnaround time?
The standard delivery time is approximately 25 working days.
How should I store lyophilized peptides?
Lyophilized peptides should be stored at -15 °C to -35 °C for long-term stability (over one year). Peptides containing Gln, Asn, or oxidation-sensitive residues (Cys, Met, Trp) may require storage at -65 °C to -85 °C. Always allow the vial to reach room temperature before opening to avoid condensation.
How can I dissolve my peptide?
Solubility depends on the peptide’s sequence and charge. Before dissolving your peptide, please read the “Guidelines for Dissolving Peptides” detailed in the Peptide Synthesis Report and perform a small-scale solubility test before dissolving the full sample.
Do I retain the rights to my peptide sequence?
Yes. Customers retain full ownership of their sequence data and all related intellectual property.
Will I lose IP or be tied to NZYtech afterward?
No. You retain IP unless a formal co-development is agreed upon. Future licensing or joint work is always case-by-case and mutually negotiated. NZYtech’s role is to enable not own your innovation.
What does the mentorship with NZYtech involve?
Each winner will be assigned a dedicated NZYtech scientific mentor for guidance, plus access to a scientific advisory group.
How often will I report progress and to whom?
You’ll participate in quarterly meetings and must submit a final report within 6 months of completing the project or using the Wallet.
Are there opportunities to co-develop a product with NZYtech?
Yes. If your project shows strong potential, you may be invited to explore co-development, validation, or licensing, subject to mutual agreement.
What type of support can I expect for manufacturing or validation?
NZYtech may provide access to internal validation tools, instruments, or manufacturing support, evaluated on a case-by-case basis.
Can awardees be featured in NZYtech press or events?
Yes. Winners may be featured on NZYtech’s website, press materials, social media, or events like MEDICA, with full GDPR compliance
Can the NZY Wallet be split across different projects or labs?
No. The Wallet is linked to the individual winner’s online account and cannot be split, transferred, or combined with other accounts.
What products and services are included in the NZYtech catalog?
The Wallet is redeemable for any NZYtech commercial products including enzymes, master mixes, lyophilization-ready reagents, diagnostic tools, and selected services.
What happens if I don’t use the full NZY Wallet within one year?
Unused credits will expire after one year. No extensions are granted, so plan usage early.
Can I combine the Wallet with institutional or national funding?
Yes. You can combine funds, provided the NZY Wallet is used specifically for purchasing NZYtech products.
Will I receive physical products or vouchers?
No. The price will be made available via the NZY Wallet and with those credits you can purchase NZYtech products exclusively.
Who will review the applications and what expertise do they bring?
A jury made up of NZYtech leadership and external experts in academia and diagnostics will evaluate all submissions.
What are the main evaluation criteria?
Scientific Excellence
Innovation Track Record
Translational Readiness
Fit with NZYtech’s platforms
Vision & Societal Impact
How do you define "translational readiness"?
It means your idea shows clear potential to move from lab to product, ideally within a 1–3 year window, and can be supported by NZYtech reagents or infrastructure.
Will early-stage ideas be considered if they have strong technical merit?
Yes, provided you show a credible path to application and how the NZY Wallet can help build a prototype or proof-of-concept.
How many finalists are selected for the pitch round?
The jury will select a shortlist of finalists (number not fixed) to present a 10-minute pitch. From this group, up to 4 winners may be chosen (1 grand prize + 1–3 excellence distinctions).
How much time will the application take?
Expect around 8–10 hours total: it requires a 250-word abstract, a 4-page proposal, a 1-page wallet usage plan, a 1-page motivation letter and your CV. But you only submit once via email.
What are the exact materials required for submission?
You must submit:
- 250-word abstract
- 4-page scientific proposal
- 1-page NZY Wallet use plan
- 1-page motivation letter
- CV
Is there a specific template for the 4-page proposal?
No fixed template, but your proposal must clearly describe objectives, methodology, and timeline. Focus on translational potential and technical soundness.
Do I need to submit in English or are Portuguese/Spanish submissions accepted?
All submissions must be in English, as the jury includes international reviewers.
What happens after I submit. When will I know if I’m selected?
Finalists will be contacted in October–November 2025 for a 10-minute pitch session. Winners will be announced in November 2025.
Can I edit my application after submitting it?
No. Once submitted, applications are considered final. Ensure all documents are complete and aligned before upload.
What can I actually buy with the NZY Wallet?
You can redeem NZY Wallet credits (€5K–€10K) for any NZYtech product. From master mixes and enzymes to lyophilization-ready reagents and diagnostic services. The prize is structured to empower you to prototype fast using the same tools NZYtech uses internally.
What counts as a strong “translational” proposal?
Winning projects must show clear short-to-mid-term potential for real-world diagnostics or biotools, especially if they integrate with NZYtech platforms (like point-of-care formats or kit modularity). Think prototype-ready, not theoretical.
Am I eligible if I’m turning 41 this year?
You are eligible as long as you are 40 years old or younger at the time of application submission, that is, by October 15th, 2025
Can I apply if I’m based in industry but collaborate with a university?
Yes, as long as you have a formal affiliation with an academic institution, hospital, startup, or R&D lab. Industry-only applicants without such ties are not eligible.
Does my project need to be diagnostic-focused, or are research tools eligible?
Both are welcome. Proposals must align with one of NZYtech’s focus areas: enzymes for life sciences, molecular diagnostics, kit development, reagent lyophilization, or point-of-care assays.
Can I apply if I’ve received other grants for the same project?
Yes, but you must clearly show how the NZY Wallet will uniquely accelerate or expand your project's translational pathway, using NZYtech reagents or services.
I work solo. Do I need a full institutional affiliation?
Yes. Affiliation with a recognized institution is mandatory. This ensures feasibility, accountability, and alignment with the prize’s collaborative intent.
Do you offer MGB Probes or alternatives?
Such type of probes have the purpose of increasing the melting temperature of the probe which can be beneficial for some assays. We offer two types of probes for customers who need such type of characteristic - MGB probes and Xs probes. These probes differ in the type of chemistry used to achieve the higher Tm.
What are the recommended storage conditions for oligonucleotides?
For best results, store oligos in a TE buffer. If chelating/buffering agents aren't suitable, opt for Low-TE buffer or sterile water.
Recommended storage order: TE > Low-TE > lyophilized/dried > water.
Ideal storage temperature is -30 °C to -15 °C. Refrigerated oligos (2 °C to 8 °C) remain stable for at least one year. Oligos stored at room temperature (dried or dissolved in TE/ Low-TE buffer are stable for at least one month.
Protect from UV light, especially for fluorophore-modified oligos, as they may bleach. Treat modified oligos like non-modified ones for optimal storage.
Is desalting included in the purification process for all standard oligo synthesis?
Yes, desalting is a standard purification procedure for all our standard oligos.
In the post-synthesis phase, during the activation of oligonucleotides for biological applications, the formation of salts occurs. Notably, these salts may serve as inhibitors, particularly in the context of polymerase chain reaction (PCR) assays.
Please be aware that specific applications, such as cloning, may necessitate or strongly benefit from additional purification.
Why does NZYtech ship oligos in TE (Tris-EDTA) buffer as the default choice?
NZYtech ships oligonucleotides in TE (Tris-EDTA) buffer as the default because it has proven to be the most stable method for storing oligos. TE buffer ensures the integrity of the DNA during transportation, preventing degradation. However, we understand that preferences may vary. If you have specific storage or shipment conditions in mind, you can choose from other standard conditions available to meet your requirements. Our goal is to provide flexibility while maintaining the high quality of our products.
What concentration options are available when ordering oligonucleotides?
By default, oligos will be shipped at a concentration of 100 µM (100 pmol/µl)
However, if the yield of an oligo intended for delivery in a 2 ml tube exceeds 180 nmol, we take a specialized approach. In such cases, we ship it at a concentration of 1 nM (1 nmol/µl). This measure is implemented to ensure the best handling and stability of your oligonucleotide order during transit.
What are the purification options offered by NZYtech, and how do they differ?
NZYtech provides multiple purification options tailored to diverse needs:
Desalting: Desalting is a standard purification procedure for all our standard oligos.
In the post-synthesis phase, during the activation of oligonucleotides for biological applications, the formation of salts occurs. Notably, these salts may serve as inhibitors, particularly in the context of polymerase chain reaction (PCR) assays.
Cartridge: A step beyond desalting, cartridge purification eliminates failure sequences, enhancing the purity of the final oligo mixture.
HPLC (High-Performance Liquid Chromatography): Ideal for achieving higher purity, HPLC removes truncated sequences and synthesis-related salts. Some modifications often require HPLC purification due to lower coupling efficiency. For certain products like dual-labelled qPCR probes, the HPLC purification step is automatically included in the price calculation as a standard feature.
PAGE (PolyAcrylamide Gel Electrophoresis): Designed to minimize impurities and ensure the highest percentage of full-length oligos. While it significantly lowers yield, PAGE purification is recommended for oligos intended for demanding applications such as mutagenesis and cloning.
What is the process for ordering dual-labelled oligos like Taqman probes from NZYtech?
NZYtech introduces a dedicated product class for these oligonucleotides, defaulting to HPLC purification and quality assessment via UPLC-MS to ensure adherence to stringent quality standards. These dual-labelled oligos are available for order under the following categories:
TDP (Twice Dyed Probes): Hydrolysis probes, serving as the equivalent of Taqman probes in qPCR.
XS probes (MGB alternative): Hydrolysis probes equivalent to MGB probes, designed to elevate the Tm for enhanced specificity.
MB probe (Molecular Beacon Probes): Featuring stem-loop structures.
FRET probe (Fluorescent Resonance Energy Transfer probes): Tailored for applications like real-time PCR.
How much oligo/probe will I receive and what are the guaranteed yield for each synthesis scale?
A: While we guarantee a minimum yield for non-labeled, non-HPLC/PAGE purified standard DNA oligos up to 30 bases, it's common for customers to receive yields surpassing this minimum guarantee:
|
Synthesis Scale |
Guaranteed Minimum Yield |
|---|---|
|
10 nmol |
10 nmol |
|
40 nmol |
20 nmol |
|
200 nmol |
95 nmol |
|
1000 nmol |
400 nmol |
The yield of an oligo depends on its sequence. Some sequences, like those with multiple consecutive dG's, may yield less, but in most cases, yields are higher.
For qPCR TDP probes, the approximate yield is:
|
Synthesis Scale |
Approximate Yield |
|---|---|
|
40 nmol |
5 nmol |
|
200 nmol |
20 nmol |
|
1000 nmol |
60 nmol |
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 ° to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
Supreme NZYProof 2x Colourless Master Mix
The master mix should be stored at -20 ºC, in a constant temperature freezer. Minimize the number of freeze-thaw cycles by storing in working aliquots. The Mix maybe stored at 4 °C for up to 7 days.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 ° to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
How should I store my NZYProof 2x Green Master Mix?
The master mix should be stored at -20 ºC, in a constant temperature freezer. Minimize the number of freeze-thaw cycles by storing in working aliquots. The Mix maybe stored at 4 °C for up to 7 days.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 ° to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
How should I store my NZYProof 2x Colourless Master Mix?
The master mix should be stored at -20 ºC, in a constant temperature freezer. Minimize the number of freeze-thaw cycles by storing in working aliquots. The Mix maybe stored at 4 °C for up to 7 days.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 ° to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Template DNA damaged or degraded. An intact, high-quality template is essential to achieve a reliable amplification of large fragments. Extreme care must be taken in the preparation and handling of DNA. Always use purified high-quality DNA as template.
d) Concentration of magnesium is too low . Mg2+ is included in the 10× Reaction Buffer at a final concentration of 2 mM, which is sufficient for most targets. For some targets, a higher Mg2+ concentration may be required. Titrate from 2 mM to 4 mM (final concentration) in 0. 5 mM increments.
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences. Check the quality and concentration of primer solutions. We recommend to prepare small-volume working aliquots from the stock solution. Avoid using primers subjected to multiple freezing-thawing cycles.
How should I store my Supreme NZYLong DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Template DNA damaged or degraded. An intact, high-quality template is essential to achieve a reliable amplification of large DNA fragments. Extreme care must be taken in the preparation and handling of DNA. Always use purified high-quality DNA as template.
d) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.5 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences. Check the quality and concentration of primer solutions. We recommend to prepare small-volume working aliquots from the stock solution. Avoid multiple freeze cycles.
How should I store my NZYLong 2× Colourless Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Template DNA damaged or degraded. An intact, high-quality template is essential to achieve a reliable amplification of large DNA fragments. Extreme care must be taken in the preparation and handling of DNA. Always use purified high-quality DNA as template.
d) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.5 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences. Check the quality and concentration of primer solutions. We recommend to prepare small-volume working aliquots from the stock solution. Avoid multiple freeze cycles.
How should I store my Supreme NZYLong 2× Colourless Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Template DNA damaged or degraded. An intact, high-quality template is essential to achieve a reliable amplification of large DNA fragments. Extreme care must be taken in the preparation and handling of DNA. Always use purified high-quality DNA as template.
d) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.5 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences. Check the quality and concentration of primer solutions. We recommend to prepare small-volume working aliquots from the stock solution. Avoid multiple freeze cycles.
How should I store my Supreme NZYLong 2× Green Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature/ primer concentration. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (3 ° to 5 °C lower than Tm). Optimise annealing temperature using temperature gradient PCR. Increase primer concentration for lower yield amplicons.
b) Inadequate cycling conditions. Increase extension time and/or number of cycles.
c) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
d) Inadequate amount of template DNA. Titrate template amount. We recommend starting with 20-50 ng of genomic DNA.
c) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.25 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another pool of primers, avoiding complementary sequences. Ensure that each primer pair originates a single product without primer-dimers in individual PCR reactions. If necessary, reduce primer concentration. Reduce the number of cycles.
What is the maximum size of amplicon using Multiplex PCR NZYTaq 2x Green Master Mix?
Multiplex PCR NZYTaq 2x Green Master Mix allows efficient multiplexing of targets ranging in size from 70 bp to 2.5 k.
Which type of DNA ends are generated in the PCR products using Multiplex PCR NZYTaq 2x Green Master Mix?
Resulting PCR products have an A-overhang and are suitable for cloning with NZYTech´s NZY-A PCR cloning kit (MB053) or NZY-A Speedy PCR cloning kit (MB137).
I don’t distinguish separate bands using agarose gel electrophoresis. What should I do?
When separating samples through agarose gel electrophoresis, please take into account that both concentration and grade of agarose are important when working with closely-sized fragments. NZYTech offers a vast portfolio of agaroses with different specifications to cover a wide range of needs. Please visit www.nzytech.com/products-services/category/molecular-biology/products/agaroses/ to identify the agarose that most suits your experiment.
How should I store my Multiplex PCR NZYTaq 2x Green Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature/ primer concentration. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (3 ° to 5 °C lower than Tm). Optimise annealing temperature using temperature gradient PCR. Increase primer concentration for lower yield amplicons.
b) Inadequate cycling conditions. Increase extension time and/or number of cycles.
c) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
d) Inadequate amount of template DNA. Titrate template amount. We recommend starting with 20-50 ng of genomic DNA.
c) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.25 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another pool of primers, avoiding complementary sequences. Ensure that each primer pair originates a single product without primer-dimers in individual PCR reactions. If necessary, reduce primer concentration. Reduce the number of cycles.
What is the maximum size of amplicon using Multiplex PCR NZYProof 2x Colourless Master Mix?
Multiplex PCR NZYColourless 2x Green Master Mix allows efficient multiplexing of targets ranging in size from 70 bp to 2.5 k.
Which type of DNA ends are generated in the PCR products using Multiplex PCR NZYProof 2x Colourless Master Mix?
Resulting PCR products have blunt-ends and are suitable for cloning with NZYTech´s NZY-blunt PCR cloning kit (MB121).
I don’t distinguish separate bands using agarose gel electrophoresis. What should I do?
When separating samples through agarose gel electrophoresis, please take into account that both concentration and grade of agarose are important when working with closely-sized fragments. NZYTech offers a vast portfolio of agaroses with different specifications to cover a wide range of needs. Please visit www.nzytech.com/products-services/category/molecular-biology/products/agaroses/ to identify the agarose that most suits your experiment.
How should I store my Multiplex PCR NZYProof 2x Colourless Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Additives required. Adding PCR-enhancing agents, like NZYTaq 5x Optimizer Solution (MB060) or NZYTaq 2x GC-Enhancer solution (MB143) may improve yield while amplifying difficult templates.
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences. Adjust concentration of magnesium. Generally, 2-3 mM MgCl2, typically 2.5 mM final concentration, works well for the majority of PCR reactions. Optimal concentration depends on target template, buffer and dNTPs. Optimise magnesium concentration by supplementing MgCl2 in 0.5 increments up to 4 mM.
How should I store my NZYTaq II DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 ° to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Inadequate PCR Setup. Adjust the primers concentration. Be sure to add the DNA template in last, and that you prepare the PCR mix at room temperature.
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences. Adjust concentration of magnesium. Generally, 2-3 mM MgCl2, typically 2.5 mM final concentration, works well for the majority of PCR reactions. Optimal concentration depends on target template, buffer and dNTPs. Optimise magnesium concentration by supplementing MgCl2 in 0.5 increments up to 4 mM.
How should I store my Supreme NZYTaq II DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.5 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences.
How should I store my Supreme NZYTaq II 2× Green Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 °C to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
c) Concentration of magnesium is too low. Mg2+ is included in the Master Mix at a final concentration of 2.5 mM, which is sufficient for most targets. For some targets, higher Mg2+ concentration may be required. Titrate from 2.5 mM to 4 mM (final concentration) in 0.5 mM increments. (Note: MgCl2 is not provided in separate tubes).
How can I reduce the number of non-specific bands?
Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences.
How should I store my NZYTaq II 2× Colourless Master Mix?
The product should be stored at -20 ºC, in a constant temperature freezer.
What to do when there is no product amplification or low yield?
This can be due to the following situations:
a) Inadequate annealing temperature. The reaction mix composition may affect the melting properties of primers and DNA. Adjust the annealing temperature to accommodate the primer with the lowest melting temperature (5 ° to 10 °C lower than Tm).
b) Presence of PCR inhibitors. Some DNA isolation procedures, particularly genomic DNA isolation, can result in the co-purification of PCR inhibitors. Reduce the volume of template DNA in reaction or dilute template DNA prior to adding to the reaction. Diluting samples even 1:10,000 has been shown to be effective in improving results, depending on initial DNA concentration.
How should I store my Speedy NZYProof DNA polymerase?
The product should be stored at -20 ºC, in a constant temperature freezer.
I’d like to clone a fragment amplified with NZYSpeedy Proof DNA polymerase. How should I proceed?
Speedy NZYProof DNA polymerase has 3´→ 5´ exonuclease activity and will generate blunt-ended products. So, blunt-end cloning is recommended, by using NZY-blunt PCR cloning kit (MB121) or other similar cloning kit. Nevertheless, if TA cloning is required, the generated blunt-ended PCR fragments can be modified by adding 3´A-overhangs with a different polymerase, such as Taq DNA polymerase (see the A-tailing protocol in the product brochure of NZY-A PCR cloning kit – MB053). Due to the strong proofreading activity of Speedy NZYProof DNA polymerase, we recommend immediately proceed with cloning after this procedure, in order to avoid lost of the 3´A-overhangs during storage. Alternatively, the PCR products can be cloned using the innovative NZYEasy Cloning & Expression Systems in a rapid, precisely and ligase-independent manner. For optimal cloning efficiencies, silica-column purification of the PCR product using NZYGelpure kit (MB011) or other similar kit is highly recommended. This procedure will remove dNTPs, unused primers and other impurities. Spin-column purification is sufficient as long as the product is >90% pure. Gel-extraction should be performed in case non-specific amplifications and/or primer-dimers are formed.
What are the benefits of this method compared to classical cloning methods?
NZYEasy Cloning & Expression System relies on a simple reaction that enables the assembly of DNA inserts into a cloning or an expression vector in a precise and predetermined order. No insert phosphorylation, blunt-end polishing or digestion is required, thus making the NZYEasy Cloning & Expression System fast when compared to classical cut and ligation methods. Furthermore, cloning can be achieved either directly using blunt-ended or Taq-generated PCR products. The cloning is performed in a single reaction with high efficiencies compared to traditional methods, even when simultaneously clone multiple inserts into one vector.
Is it necessary to purify PCR products?
Spin-column purification of PCR products is highly recommended to remove dNTPs and impurities resulting from the amplification. Presence of non-specific amplification products and/or primer-dimers requires gel-purification of the amplified nucleic acid, which will enhance cloning efficiencies.
To amplify the insert, do I need to use PCR primers that have been purified by PAGE or HPLC?
No, you can use standard, desalted primers.
The insert will be cloned in reading-frame into pHTP vectors?
Yes. Inserts correctly cloned into pHTP1 expression vector will maintain reading frames starting on the ATG codon.
What type of competent cells is suitable to transform DNA clones obtained from the ligase-independent cloning reaction?
We recommend using E. coli DH5α (NZYTech, cat. No. MB004) as a host for cloning. Regarding expression, all E. coli strains expressing the gene that codifies for T7 RNA polymerase, such as BL21(DE3) (NZYTech, cat. No. MB006), are suitable to use.
It is possible to clone many different inserts into one vector?
Yes, the NZYEasy Cloning & Expression System allows multiple fragment cloning into one pHTP vector. DNA fragments will be correctly annealed together in the desired order on the pHTP vector through the action of the highly efficient NZYEasy enzyme mix, free of any DNA ligase. Just combine the PCR-generated fragments with appropriate complementary overhangs, by allowing assembling in the order you choose, and a linearized pHTP vector. Both first and last inserts should have overhangs complementary with respective ends of the vector, thereby allowing efficient cloning. Each DNA fragment, encoding for an individual domain of the final multidomain construct, are kept in frame and are separated from each other by a linker sequence of 6 amino acids.
How many DNA fragments can be cloned in one reaction?
The number of inserts that can be joined in the same vector depends on their length and sequence. NZYEasy Cloning & Expression System has been tested efficiently for cloning 5 different genes of around 500-700 bp in size in the same vector. Note that the cloning efficiency decreases as the size of the final construct increases. However, if the simultaneous cloning of the desired inserts does not work, you can try a sequential cloning approach.
How can I reduce the number of false positive colonies containing the vector-backbone only?
Gel-purify the PCR band of interest in order to remove the template plasmid. Alternatively, digest PCR products with DpnI (NZYTech cat No. MB078) restriction enzyme to eliminate parental methylated DNA templates. When possible, you can also reduce the amount of template plasmid DNA to 0.1-0.5 ng per 50 µL PCR reaction.
The cloning reactions originate no colonies or a few number of colonies. What should I do?
Perform a cloning reaction using the PCR fragment provided as positive control with NZYEasy Cloning & Expression kits. Using this procedure, you can evaluate the functionality of cloning and transformation conditions.
If cloning and transformation result successfully using the positive control, then make sure that your DNA insert and the linearized pHTP vector share the required end-terminal complementarity by analysing primer sequences; verify the quality of the insert by gel electrophoresis; optimize the ligase-independent cloning reaction trying different vector:insert molar ratio, and/or repeat PCR and gel-purify the PCR product for a new cloning reaction and transformation. Alternatively, consider that the cloned insert may be toxic to E. coli. If this is the case, and if you are directly cloning into the expression vector, try to clone first into the low-copy number pHTP0 vector.
If cloning and transformation do not result successfully, check also the transformation efficiency of E. coli competent cells; verify if LB plates contain the appropriate antibiotic for the pHTP vector you are using; make sure you correctly handle the NZYEasy enzyme mix; verify if a specific component is missing in the cloning reaction.
Why should I use NZYTech’s gene synthesis services?
– You can focus on your research while we produce your gene
– It is possible to codon optimize your gene, thus increasing the possibility of achieving high protein levels
– Gene synthesis provides access to DNA templates that are not easily available on nature
Is it possible to synthesize sequences with high/low GC content or repeats?
Yes, NZYtech successfully synthesizes complex genes. However, sequences containing several repeated sequences and regions with very high/low GC content may not be accepted for synthesis.
Is it possible to optimize gene design for high levels of expression?
Yes, ATGenium allows optimizing not only critical factors in transcription and translation but also factors related with co-translational protein folding. ATGenium comprises an extensive collection of codon usage tables, allowing optimizing codon usage for improved protein expression in different host systems.
Are all genes sequence-verified?
All genes are 100% sequence verified.
In which vector am I going to receive my gene?
Synthetic genes can be cloned into our standard cloning vectors (pUC57), into pHTP Escherichia coli expression vectors or into a selection of other bacterial, mammalian and viral expression vectors. You can find the complete list of available vectors on “Description” section.
How much plasmid DNA do you deliver?
NZYtech delivers ≥1 µg of plasmid DNA (standard quantification by absorbance-based methods). For more accurate and specific DNA quantification, fluorescence-based methods are available upon request (additional charge applies).
What is NZYTech’s policy for Intellectual Property Protection?
NZYTech does not claim rights or ownership of any intellectual property related to the DNA/amino acid sequences provided by our customers or the resulting synthetic genes. No data or material will be released to a third party without your consent. All genes resulting from gene synthesis and/or cloning services are stored at NZYTech for a three months period. After this period nucleic acids provided to customers are discarded.
Why should I use NZYtech’s gene synthesis services?
– You can focus on your research while we produce your gene
– It is possible to codon optimize your gene, thus increasing the possibility of achieving high protein levels
– Gene synthesis provides access to DNA templates that are not easily available on nature
Is it possible to synthesize sequences with high/low GC content or repeats?
Yes, NZYtech successfully synthesizes complex genes. However, sequences containing several repeated sequences and regions with very high/low GC content may not be accepted for synthesis.
Is it possible to optimize gene design for high levels of expression?
Yes, ATGenium allows optimizing not only critical factors in transcription and translation but also factors related with co-translational protein folding. ATGenium comprises an extensive collection of codon usage tables, allowing optimizing codon usage for improved protein expression in different host systems.
Are all genes sequence-verified?
All genes are 100% sequence verified.
In which vector am I going to receive my gene?
Synthetic genes can be cloned into our standard cloning vectors (pUC57), into pHTP Escherichia coli expression vectors or into a selection of other bacterial, mammalian, yeast and baculovirus/insect expression vectors. You can find the complete list of these additional vectors in "Additional Expression Vectors List" in the "Manuals" section.
How much plasmid DNA do you deliver?
NZYtech delivers ≥1 µg of plasmid DNA.
What is NZYtech’s policy for Intellectual Property Protection?
NZYtech does not claim rights or ownership of any intellectual property related to the DNA/amino acid sequences provided by our customers or the resulting synthetic genes. No data or material will be released to a third party without your consent. All genes resulting from gene synthesis and/or cloning services are stored at NZYTech for a three months period. After this period nucleic acids provided to customers are discarded.
Is it possible to optimize gene design for high levels of expression?
Yes, ATGenium allows optimizing not only critical factors in transcription and translation but also factors related with co-translational protein folding. ATGenium comprises an extensive collection of codon usage tables, allowing optimizing codon usage for improved protein expression in different host systems.
Why should I use NZYtech’s gene synthesis services?
– You can focus on your research while we produce your gene
– It is possible to codon optimize your gene, thus increasing the possibility of achieving high protein levels, and increasing the possibility of obtaining a non-complex sequence (compatible with Gene Fragments synthesis)
– Gene synthesis provides access to DNA templates that are not easily available on nature
Is it possible to synthesize gene fragments with high/low GC content or repeats?
Yes, NZYtech successfully synthesizes complex gene fragments. However, sequences containing several repeated sequences and regions with very high/low GC content may not be accepted for synthesis.
Are Gene Fragments sequence-verified?
No, Gene Fragments quality control procedure only involves size verification by electrophoresis. NZYtech’s Gene Fragments are synthesized under the same careful conditions as our custom gene synthesis service. However, since Gene Fragments are delivered to you as uncloned products the existence of minor populations of mutated or truncated fragments is inevitable.
In which vector am I going to receive my gene fragment?
Gene fragments are delivered as uncloned products.
How do you deliver synthetic gene fragments?
NZYtech standardly delivers ≥400 ng dry dsDNA fragments. Please note that due to inherent losses during the freeze-drying process post-aliquoting, the final yield may vary slightly. To compensate for this and ensure sufficient plasmid quantity, a slightly higher amount than specified is typically provided. We recommend resuspending the sample upon receipt and performing a new concentration measurement prior to use.
For higher accuracy and specificity, fluorescence-based quantification of dsDNA is available upon request (additional charges apply). In this case, the dsDNA is delivered in liquid form, resuspended in water.
What is NZYtech’s policy for Intellectual Property Protection?
NZYtech does not claim rights or ownership of any intellectual property related to the DNA/amino acid sequences provided by our customers or the resulting synthetic genes. No data or material will be released to a third party without your consent. Gene fragments produced are not stored in-house after dispatch to clients.
What payment methods can I use?
You can use Credit Card (Visa, MasterCard), PayPal and Bank Transfer
How long will it take to receive my order?
For most countries, the delivery occurs within five working days, except in case of stock rupture. However, it can take longer depending on the destination of the order.
Do we deliver our products on weekends?
All orders are delivered during the work week.
How much does the shipping cost?
Portugal (Main Land): 16 €
Spain (Main Land): 20€
All other geographic areas: Upon quotation (in these cases shipping costs are additional to the price of the products and fully supported by the client).
Products that require ultra-low temperature shipping will have an extra cost for dry ice (39 € / shipment for EU members + UK + Switzerland and 290€ / shipment for all other countries).
Products that require Dangerous Goods Handling Charges will have an extra cost for (20 € / shipment for Portugal and Spain - 100 € for all other countries).
Custom fees, if any, related to the clearance of goods upon product arrival to the destiny country are of CUSTOMER responsibility.
What is your return policy?
NZYTech CUSTOMER Service is available to assist you in every way possible if a problem arises with your order. The CUSTOMER must inspect all packages immediately upon receipt and store the goods at the designated storage temperature.
CUSTOMER will only have 14 calendar days to notify NZYTech if any damage or discrepancy occurs. After this period, NZYTech will assume that the products were properly delivered and no longer accepts any complaint regarding packaging, delivery or other. In case of any damage occur, please retain the damaged goods, packing material and shipping documents and please notify our Customer Service through e-mail at info@nzytech.com. If the damage requires you to return the goods to NZYTech, our Customer Service will provide assistance, shipping instructions and authorizations. If an item is shipped to you incorrectly, as the result of a mistake from our Logistics team, please contact our Customer Service through e-mail at info@nzytech.com, and we will take a quick and appropriate action to correct the problem. At NZYTech sole discretion, returns may be accepted for items stored at room temperature and that have been ordered by mistake but will be subjected to a processing fee of 30% to cover costs.
For e-commerce online orders carried out by consumers, if applicable, the return period without any cause is 14 calendar days from date of receipt of goods.
What is my warranty?
NZYTech warrants that its products comply with the specifications indicated in the accompanying technical brochure as long as the product itself has a valid Certificate of Analysis (CoA). If a product fails to comply with its specifications, NZYTech should be informed as it may choose to replace it free of charge or refund the purchase price. This warranty applies only to products that have been handled and stored in accordance with NZYT recommendations/specifications.
Are all job opportunities listed on the website?
Yes, all currently active employment opportunities are listed on this website. Please refer to the Career section.
What is the best way to apply for a position?
It is strongly recommend that you apply using the online form available on this page.
I applied for an available position. What are the next steps?
When you apply on-line for a specific position, you will receive a confirmation via email, and the information you provided becomes available to our Human Resources Department. Qualified applicants will then be contacted by our Human Resources representative or the hiring manager
Can we request samples?
NZYTech can provide samples after the definition of customer needs and feasibility of product is confirmed.
Does NZYTech offer lyophilized products?
Yes, NZYTech can provide a broad range of lyo-ready or lyophilized product, both off-the-shelf selection or for a customized formulations
Are there additional fees, such as licensing?
NZYTech does not add any licensing fees. Quotes are provided as a one-off total sum and are note dependent on continued use after delivery.