A simple and fast method for cloning and analyzing polymerase chain reaction products.
A method describing a fast and efficient way for cloning polymerase chain reaction (PCR) products is presented that involves end repair and purification of the PCR product, followed by kinasing and ligation to the vector with the use of a temperature gradient. Efficiency of ligation was estimated to be 50%-70%. Following transformation, cells are plated on MacConkey agar. Bacteria from selected colonies are used directly from the plates for screening without any subsequent purification. Using this protocol, PCR products can be efficiently cloned quickly and economically.
Skryabin
B
B
Fishberg Center for Neurobiology, Mount Sinai School of Medicine, New York City, New York 10029.
Vassilacopoulou
D
D
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D004260:DNA Repair; D008967:Molecular Biology / Q000191:economics / Q000379:methods; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012641:Selection, Genetic; D014170:Transformation, Genetic