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

Y. Piao, N. T. Ko, M. K. Lim, Minoru Ko

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

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 ∼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.

Original languageEnglish
Pages (from-to)1553-1558
Number of pages6
JournalGenome Research
Volume11
Issue number9
DOIs
Publication statusPublished - 2001
Externally publishedYes

Fingerprint

Gene Library
Libraries
RNA
Polymerase Chain Reaction
Initiator Codon
DNA
Blastocyst
Complex Mixtures
Complementary DNA
Clone Cells
Kidney

ASJC Scopus subject areas

  • Genetics

Cite this

Construction of long-transcript enriched cDNA libraries from submicrogram amounts of total RNAs by a universal PCR amplification method. / Piao, Y.; Ko, N. T.; Lim, M. K.; Ko, Minoru.

In: Genome Research, Vol. 11, No. 9, 2001, p. 1553-1558.

Research output: Contribution to journalArticle

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