NZY-A Speedy PCR cloning kit (MB137)

Refrigerated package

155,00207,00

Description: NZY-A Speedy PCR cloning kit was designed to carry out fast and efficient cloning of PCR products with 3´-A overhangs, which result from amplifications using non-proofreading DNA polymerases. This methodology combines the efficiency of an improved ligation buffer with the speed of Speedy Ligase to allow a rapid ligation between the vector and the PCR product in only 5 minutes at room temperature (18-25 °C). The cloning vector was prepared by cutting NZYTech´s pNZY28 with EcoR V and adding a 3´ terminal thymidine at both ends. These single 3´-T overhangs improve the efficiency of ligation of a PCR product into plasmids by preventing recircularization of the vector and providing compatible overhangs for PCR products generated by non-proofreading thermostable polymerases. Vector pNZY28 contains multiple restriction sites within the multiple cloning region. However, vector digestion with EcoR I or BamH I allows the release of the PCR product since the vector cloning region is flanked by recognition sites of both enzymes.

Features:
– Fast and easy TA cloning procedure
– High-efficiency cloning (>90% of positive clones)
– No requirement for time-consuming  restriction digests
– Possibility of blue/white screening

Applications:
– Cloning of A overhangs PCR products
– Possibility of clone blunt-ended PCR products after an A-tailing procedure

Specifications:

Cloning method: TA PCR
Ligation reaction time: 5 min
Ligation reaction temperature: room temperature
Storage conditions: Store competent cells at -80 °C. Other kit components may be stored at -20 °C or at -80 °C
Shipping conditions: Shipped at dry ice

Components:
– NZY-A Speedy Buffer (4x)
– pNZY28-A vector (50 ng/µL)
– Speedy Ligase (5 U/µL)
– NZY-A positive control insert (50 ng/µL)
– NZYStar Competent Cells (*)
– Competent Cells Control Plasmid (0.1 ng/µL) (*)

(*) only provided in MB13701 kit

pNZY28 vector map
Product Brochure EN
Safety Information EN
Informacao de Seguranca PT
Certificate of Analysis YO031

1. Schematic representation of the NZY-A Speedy PCR cloning method

Scheme - NZY-A Speedy PCR cloning kit

2. Protocol Overview:

TACloning_Protocol

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