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XB-ART-26119
Genet Anal Tech Appl 1990 Feb 01;71:5-17. doi: 10.1016/0735-0651(90)90038-h.
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Cloning of PCR-amplified total cDNA: construction of a mouse oocyte cDNA library.

Welsh J , Liu JP , Efstratiadis A .


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.

PubMed ID: 1691011
Article link: Genet Anal Tech Appl
Grant support: [+]