Screening recombinant DNA libraries: a rapid and efficient method for isolating cDNA clones utilizing the PCR.
We describe an expeditious method for the isolation of cDNA clones utilizing PCR-based amplification of target sequences from cDNA libraries. This method is rapid, less labor-intensive and inexpensive when compared with screening libraries with radiolabeled probes. This method can be applied to isolate multiple members of a protein family as well as homologous genes in different species by designing appropriate primers to amplify the most conserved regions. Utilizing this method, a novel reticulocyte CD44 transcript was isolated.
Isola
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University of Pittsburgh School of Medicine, Department of Pathology, PA 15261.
Harn
H J
HJ
Cooper
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DL
D010582:Bacteriophage lambda; D001483:Base Sequence; D004247:DNA / Q000302:isolation & purification*; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D016201:Receptors, Lymphocyte Homing / Q000235:genetics
D015345:Oligonucleotide Probes; D016201:Receptors, Lymphocyte Homing; D004247:DNA