Arbitrarily primed PCR fingerprinting of RNA.
Fingerprinting of RNA populations was achieved using an arbitrarily selected primer at low stringency for first and second strand cDNA synthesis. PCR amplification was then used to amplify the products. The method required only a few nanograms of total RNA and was unaffected by low levels of genomic double stranded DNA contamination. A reproducible pattern of ten to twenty clearly visible PCR products was obtained from any one tissue. Differences in PCR fingerprints were detected for RNAs from the same tissue isolated from different mouse strains and for RNAs from different tissues from the same mouse. The strain-specific differences revealed are probably due to sequence polymorphisms and should be useful for genetic mapping of genes. The tissue-specific differences revealed may be useful for studying differential gene expression. Examples of tissue-specific differences were cloned. Differential expression was confirmed for these products by Northern analysis and DNA sequencing uncovered two new tissue-specific messages. The method should be applicable to the detection of differences between RNA populations in a wide variety of situations.
Welsh
J
J
California Institute of Biological Research, La Jolla 92037.
Chada
K
K
Dalal
S S
SS
Cheng
R
R
Ralph
D
D
McClelland
M
M
D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009928:Organ Specificity; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic; D012313:RNA / Q000235:genetics*; D012194:RNA-Directed DNA Polymerase
D009838:Oligodeoxyribonucleotides; D012313:RNA; D004247:DNA; D012194:RNA-Directed DNA Polymerase