pPCV, a versatile vector for cloning PCR products.
The efficiency of PCR product cloning depends on the nature of the DNA polymerase employed because amplicons may have blunt-ends or 3' adenosines overhangs. Therefore, for amplicon cloning, available commercial vectors are either blunt-ended or have a single 3' overhanging thymidine. The aim of this work was to offer in a single vector the ability to clone both types of PCR products. For that purpose, a minimal polylinker was designed to include restriction sites for EcoRV and XcmI which enable direct cloning of amplicons bearing blunt-ends or A-overhangs, respectively, still offering blue/white selection. When tested, the resulting vector, pPCV, presented high efficiency cloning of both types of amplicons.
Centro de Biotecnologia Molecular, Departamento de Biologia Celular, Universidade de Brasília, Brasília, DF, 70910-900 Brazil.
Christiane R
CR
Janner
Ana Lívia P
AL
Brito
Lidia Maria P
LM
Moraes
Viviane Cb
VC
Reis
Fernando Ag
FA
Torres
Molecular cloning; Plasmid; Polymerase chain reaction