The generation of radiolabeled DNA and RNA probes with polymerase chain reaction.
作者： David B. Schowalter , Steve S. Sommer
DOI: 10.1016/0003-2697(89)90019-5
关键词:
摘要: By including a radioactive triphosphate during polymerase chain reaction (PCR), probes of very high specific activity can be generated. The advantages PCR labeling include (1) uniform with 5 X 10(9) cpm/micrograms or higher (sensitivity detection: 0.028 pg target DNA per 24 h); (2) ease regulation both the and amount labeled probe produced; (3) efficient fragments less than 500 bp; (4) incorporation over wide range input template; (5) subnanogram amounts DNA; (6) direct genomic DNA. minimal allows virtually unlimited number reactions to performed on generated by one amplification under previously described nonlabeling conditions. This obviates need for CsCl gradients other large scale methods preparation. above except also achieved transcript after an where primers contain phage promoter sequence.
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