Construction of long-transcript enriched cDNA libraries from submicrogram amounts of total RNAs by a universal PCR amplification method

被引：22

作者：

Piao, Y ^{[1
]}

Ko, NT ^{[1
]}

Lim, MK ^{[1
]}

Ko, MSH ^{[1
]}

机构：

[1] NIA, Genet Lab, Dev Genom & Aging Sect, NIH, Baltimore, MD 21224 USA

来源：

GENOME RESEARCH | 2001年 / 11卷 / 09期

关键词：

D O I：

10.1101/gr.185501

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Here we report a novel design of linker primer that allows one to differentially amplify long tracts (average 3.0 kb with size ranges of 1-7 kb) or short DNAs (average 1.5 kb with size ranges of 0.5-3 kb) from a complex mixture. The method allows one to generate cDNA libraries enriched for long transcripts without size selection of insert DNAs. One representative library from newborn kidney includes 70% of clones bearing ATG start codons. A comparable library has been generated from 20 mouse blastocysts, containing only similar to 40 ng of total RNA. This universal PCR amplification scheme can provide a route to isolate very large cDNAs, even if they are expressed at very low levels.

引用

页码：1553 / 1558

页数：6

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