A simple and eficient method for high fidelity PCR cloning using antibody-neutralizing technology.
We introduce the TA cloning antibody method for the high-fidelity PCR product amplified by family B DNA polymerase without purification. This method uses antibodies and Thermus aquaticus (Taq) DNA polymerase. The antibodies can inhibit only the activity of family B DNA polymerase, and Taq can co-work for A-tailing. This method has nearly cloning efficiency to that of the PCR product of Taq.
Kitabayashi
Masao
M
Graduate School of Biosphere Sciences, Hiroshima University, Kagamiyama, Higashi-Hiroshima, Japan.
Nishiya
Yoshiaki
Y
Esaka
Muneharu
M
D000907:Antibodies, Bacterial / Q000737:chemistry*; D003001:Cloning, Molecular / Q000379:methods*; D004259:DNA-Directed DNA Polymerase / Q000276:immunology; D005822:Genetic Vectors / Q000235:genetics; D009500:Neutralization Tests / Q000379:methods; D019384:Nucleic Acid Synthesis Inhibitors; D016133:Polymerase Chain Reaction / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism; D013824:Thermus / Q000201:enzymology / Q000235:genetics
D000907:Antibodies, Bacterial; D019384:Nucleic Acid Synthesis Inhibitors; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase