CLONING OF PCR-AMPLIFIED TOTAL CDNA - CONSTRUCTION OF A MOUSE OOCYTE CDNA LIBRARY

被引：39

作者：

WELSH, J ^{[1
]}

LIU, JP ^{[1
]}

EFSTRATIADIS, A ^{[1
]}

机构：

[1] COLUMBIA UNIV,DEPT GENET & DEV,701 W 168TH ST,NEW YORK,NY 10032

来源：

GENETIC ANALYSIS-BIOMOLECULAR ENGINEERING | 1990年 / 7卷 / 01期

关键词：

D O I：

10.1016/0735-0651(90)90038-H

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. © 1990.

引用

页码：5 / 17

页数：13

共 63 条

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