InsTAclone PCR Cloning Kit

Catalog number: K1213

Thermo Scientific™  Related applications: PCR Cloning

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Description

Thermo Scientific InsTAclone PCR Cloning Kit, sufficient for 10 reactions, is a TA system for direct one-step cloning of PCR products with 3'-dA overhangs generated by Taq DNA polymerase and other thermostable DNA polymerases which lack proofreading activity. The high quality ready-to-use TA cloning vector pTZ57R/T (see Figure 1 in Supporting Data) was prepared by linearizing the pTZ57R plasmid with Eco32I and tailing with single ddT. The 3'-ddT overhangs at both ends of the cloning site prevent recircularization of the vector during ligation, resulting in high cloning yields and low background.

To increase the speed, convenience, and efficiency of cloning experiment, the kit has been combined with the unique TransformAid Bacterial Transformation Kit—a set of solutions for preparation of chemically competent E. coli cells.

Highlights

Fast cloning—approximately one hour from PCR completion to cell plating
High efficiency—more than 90% of the recombinant clones contain the target DNA
One-step procedure—additional modifications of PCR fragment are not required
Compatibility—compatible with Taq, Tth, Tfl, and other non-proofreading DNA polymerases as well as with Long PCR Enzyme Mix
Convenience of pTZ57R/T cloning vector:

• Ready-to-use: linearized with Eco32I and 3'-ddT tailed
• MCS designed for easy mapping and manipulation of the cloned insert
• Blue/white screening
• M13/pUC primer sites for sequencing or colony PCR screening
• T7 promoter for in vitro transcription of the insert

According to the protocol, ligation and preparation of competent cells is performed in parallel. Therefore, it takes only one hour from the completion of PCR to cell plating. Our transformation protocol is often faster than the transformation of commercially available competent cells. The DNA insert can be readily excised from the versatile polylinker of pTZ57R/T, sequenced using standard M13/pUC primers or in vitro transcribed with T7 RNA polymerase.

Applications

• TA cloning
• Sequencing of cloned insert
in vitro transcription of insert DNA

Related Products
InsTAclone PCR Cloning Kit
For Research Use Only. Not for use in diagnostic procedures.

Specifications

Vector Type: pTZ57R/T
Cloning Method: TA Cloning®
Bacterial or Yeast Strain: Not Included
Antibiotic Resistance (Bacterial): Ampicillin (AmpR)
Product Size: 10 reactions

Documents

Manuals & protocols

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