Literature DB >> 1691011

Cloning of PCR-amplified total cDNA: construction of a mouse oocyte cDNA library.

J Welsh1, J P Liu, A Efstratiadis.   

Abstract

We describe a general method for the synthesis and cloning of cDNA, applicable to cases in which the availability of biological material for mRNA extraction is extremely limited. A protocol allowing amplification of a heterogeneous mixture of cDNAs by the polymerase chain reaction has been devised and applied successfully to the construction of an apparently representative cDNA library, using as a model of a scarce RNA source 50 mouse ovulated eggs that can yield a maximum of 1.75 ng of poly(A)+ RNA. However, about 5% of the material obtained after amplification was adequate for cloning. Using the cloned sequences, we have derived a preliminary indirect measurement of the sequence complexity of the maternal poly(A)+ RNA in this mammalian oocyte.

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Year:  1990        PMID: 1691011     DOI: 10.1016/0735-0651(90)90038-h

Source DB:  PubMed          Journal:  Genet Anal Tech Appl        ISSN: 1050-3862


  6 in total

1.  Arbitrarily primed PCR fingerprinting of RNA.

Authors:  J Welsh; K Chada; S S Dalal; R Cheng; D Ralph; M McClelland
Journal:  Nucleic Acids Res       Date:  1992-10-11       Impact factor: 16.971

2.  Rapid isolation of desired sequences from lone linker PCR amplified cDNA mixtures: application to identification and recovery of expressed sequences in cloned genomic DNA.

Authors:  K Abe
Journal:  Mamm Genome       Date:  1992       Impact factor: 2.957

3.  A 'one tube reaction' for synthesis and amplification of total cDNA from small numbers of cells.

Authors:  R H Don; P T Cox; J S Mattick
Journal:  Nucleic Acids Res       Date:  1993-02-11       Impact factor: 16.971

4.  Serological differentiation between cystic and alveolar echinococcosis by use of recombinant larval antigens.

Authors:  M Helbig; P Frosch; P Kern; M Frosch
Journal:  J Clin Microbiol       Date:  1993-12       Impact factor: 5.948

5.  Construction and characterization of a normalized cDNA library.

Authors:  M B Soares; M F Bonaldo; P Jelene; L Su; L Lawton; A Efstratiadis
Journal:  Proc Natl Acad Sci U S A       Date:  1994-09-27       Impact factor: 11.205

6.  Molecular cloning of a serine proteinase inhibitor from Brugia malayi.

Authors:  P Yenbutr; A L Scott
Journal:  Infect Immun       Date:  1995-05       Impact factor: 3.441

  6 in total

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