PCR to identify specific clones of interest for DNA sequencing.
We describe a rapid method for identifying specific clones of interest for the purpose of sequencing. The method essentially is polymerase chain reaction using one internal primer and one vector specific primer. The procedure is particularly useful when relatively large numbers of clones are to be examined either to establish the nucleotide sequence of a full-length cDNA or to find a specific section of a large DNA. The relative orientations of inserts in different clones can also be determined using the same procedure.
Sinha
D
D
Department of Biochemistry, Temple University School of Medicine, Philadelphia, PA 19140.
Bakhshi
M R
MR
Yang
X
X
Kirby
E P
EP
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006801:Humans; D007202:Indicators and Reagents; D008099:Liver / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D000515:alpha 1-Antitrypsin / Q000235:genetics*
D007202:Indicators and Reagents; D000515:alpha 1-Antitrypsin; D004247:DNA