InsTAclone PCR Cloning Kit|現(xiàn)貨
說(shuō)明：
The InsTAclone™ PCR Cloning Kit is a TA system for direct one-step cloning of PCR products with 3'-dA overhangs (1) 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 Fig 1) 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 InsTAclone™ PCR Cloning Kit has been combined with the unique TransformAid™ Bacterial Transformation Kit – a set of solutions for preparation of chemically competent E. coli cells. 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.
成分：
Vector pTZ57R/T (pTZ57R DNA sequence in txt format)
T4 DNA Ligase
5X Ligation Buffer
Control PCR Fragment (sequence of control PCR product in txt format)
Control DNA 1
Control DNA 2
Water, nuclease-free
TransformAid™ Bacterial Transformation System:
C-Medium;
T-Solution (A);
T-Solution (B)
Detailed Protocol
質(zhì)量控制：
The kit is functionally tested by the cloning efficiency of a control PCR product. Typical yield of the recombinant clones is higher than 90%. The transformation system - TransformAid™ Bacterial Transformation Kit is tested in transformation of E.coli strains XL1-Blue and JM107 with pUC19 DNA. Typical transformation efficiency is more than 107 transformants per µg of supercoiled pUC19 DNA.
應(yīng)用：
TA cloning.
Sequencing of cloned insert.
in vitro transcription of insert DNA.