ARBITRARILY PRIMED PCR FINGERPRINTING OF RNA
ARBITRARILY PRIMED PCR FINGERPRINTING OF RNA
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DOI：
10.1093/nar/20.19.4965
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发表时间：
1992-10-11
影响因子：
14.9
通讯作者：
MCCLELLAND, M
中科院分区：
文献类型：
--
作者：
WELSH, J;CHADA, K;MCCLELLAND, M
使用任意选择的引物以低严格性对第一和第二链cDNA合成进行RNA群体的指纹分析。然后使用PCR扩增来扩增产物。该方法仅需要几纳克的总RNA，并且不受低水平的基因组双链DNA污染的影响。从任何一种组织中获得10至20个清晰可见的PCR产物的可重复模式。检测来自不同小鼠品系的相同组织的RNA和来自相同小鼠的不同组织的RNA的PCR指纹的差异。菌株特异性差异的揭示可能是由于序列多态性，应该是有用的基因的遗传作图。揭示的组织特异性差异可能有助于研究差异基因表达。克隆了组织特异性差异的实例。通过北方分析和DNA测序证实了这些产物的差异表达，发现了两个新的组织特异性信息。该方法应适用于在各种情况下检测RNA群体之间的差异。
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.