FAQ

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.

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 M_gCl₂, 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 M_gCl₂ in 0.5 increments up to 4 mM.

The product should be stored at -20 ºC, in a constant temperature freezer.

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.

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.

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. 

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.

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.

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.

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. 

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:

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:

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.

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.

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.

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.

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.

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.

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.

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.

The product should be stored at -20 ºC, in a constant temperature freezer.

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).

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.

The product should be stored at -20 ºC, in a constant temperature freezer.

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).

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.

 The product should be stored at -20 ºC, in a constant temperature freezer.

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).

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.

 The product should be stored at -20 ºC, in a constant temperature freezer.

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 T_m). 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).

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.

 Multiplex PCR NZYTaq 2x Green Master Mix allows efficient multiplexing of targets ranging in size from 70 bp to 2.5 k.

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).

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.

The product should be stored at -20 ºC, in a constant temperature freezer.

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 T_m). 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).

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.

Multiplex PCR NZYTaq 2x Colourless Master Mix allows efficient multiplexing of targets ranging in size from 70 bp to 2.5 k.

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).

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.

The product should be stored at -20 ºC, in a constant temperature freezer.

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 T_m). 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: M_gCl₂ is not provided in separate tubes).

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.

Multiplex PCR NZYColourless 2x Green Master Mix allows efficient multiplexing of targets ranging in size from 70 bp to 2.5 k.

Resulting PCR products have blunt-ends and are suitable for cloning with NZYTech´s NZY-blunt PCR cloning kit (MB121).

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.

The product should be stored at -20 ºC, in a constant temperature freezer.

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.

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 M_gCl₂, 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 M_gCl₂ in 0.5 increments up to 4 mM.

 The product should be stored at -20 ºC, in a constant temperature freezer.

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: M_gCl₂ is not provided in separate tubes).

Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences.

The product should be stored at -20 ºC, in a constant temperature freezer.

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).

Adjust annealing conditions and/or design another set of primers, by increasing the length and avoiding complementary sequences.

The product should be stored at -20 ºC, in a constant temperature freezer.

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.

 The product should be stored at -20 ºC, in a constant temperature freezer.

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.

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.

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.

No, you can use standard, desalted primers.

Yes. Inserts correctly cloned into pHTP1 expression vector will maintain reading frames starting on the ATG codon.

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.

No. pHTP vectors were designed to contain the required insert-complementary overhangs generated by PCR, as well as the specific sequence recognized by the NZYEasy enzyme mix. In theory, any vector can be converted into a “pHTP vector” to function with the NZYEasy Cloning & Expression System. In case vectors available in our pHTP portfolio do not fit your needs, please contact us at services@nzytech.com. Solubility tags can easily be included in the pHTP1 backbone and other vectors can be engineered to be used with this system. The NZYTech R&D team is available to assist you in case of need.

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.

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.

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.

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.

– 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

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.

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.

All genes are 100% sequence verified.

Synthetic genes can be cloned into our standard cloning vectors (pNZY30), 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.

NZYtech delivers 40 ng to 4 µg of plasmid DNA.

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.

– 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

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.

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.

All genes are 100% sequence verified.

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.

NZYtech delivers 40 ng to 4 µg of plasmid DNA.

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.

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.

– 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

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.

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. 

 Gene fragments are delivered as uncloned products.

NZYtech delivers ~400 ng dry dsDNA fragments.

 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.

You can use Credit Card (Visa, MasterCard), PayPal and Bank Transfer

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.

All orders are delivered during the work week.

 
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.

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.

 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.

Yes, all currently active employment opportunities are listed on this website. Please refer to the Career section.

It is strongly recommend that you apply using the online form available on this page.

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

NZYTech can provide samples after the definition of customer needs and feasibility of product is confirmed.

Yes, NZYTech can provide a broad range of lyo-ready or lyophilized product, both off-the-shelf selection or for a customized formulations

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.