The Thermo Scientific InsTAclone PCR Cloning Kit is a TA system for direct one-step cloning
of PCR products with 3’-dA overhangs (1). The high quality TA cloning vector pTZ57R/T is
ready to use for efficient ligation with PCR products providing high cloning yields and low
background. To increase the speed, convenience and efficiency of cloning, the InsTAclone™
PCR Cloning Kit has been combined with the TransformAid™ Bacterial Transformation Kit – a
set of solutions for preparation of chemically competent E. coli cells. According to our protocol,
ligation and preparation of competent cells is performed in parallel. Therefore, it only takes
approximately one hour from the completion of PCR to plating of transformed cells. Our
transformation protocol is often faster than 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.