Specific DNA amplification utilizing the polymerase chain reaction and random oligonucleotide primers: application to the analysis of antigen receptor variable regions.
The polymerase chain reaction (PCR) allows rapid amplification of DNA of known sequence. In many situations, part of a genetic sequence is known, but adjacent sequences of interest are unknown. This is common in investigations of antigen receptor genes from B and T lymphocytes, which are composed of a constant region of known sequence and a variable region, for which the sequence may not be known. Herein is described a method to amplify DNA when sequence information is available for only one primer. This procedure utilizes a primer of known sequence in conjunction with a mixture of short random primers. Application of this method to the amplification of T-cell antigen receptor cDNA is described.
Williams
W V
WV
Rheumatology Section, University of Pennsylvania School of Medicine, Philadelphia 19104.
Sato
A
A
Rossman
M
M
Fang
Q
Q
Weiner
D B
DB
D001172:Arthritis, Rheumatoid / Q000276:immunology; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004277:DNA, Single-Stranded; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*; D013582:Synovial Fluid / Q000166:cytology; D013601:T-Lymphocytes / Q000378:metabolism; D013698:Templates, Genetic
D004277:DNA, Single-Stranded; D009838:Oligodeoxyribonucleotides; D011948:Receptors, Antigen, T-Cell