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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 29, No. 4, 2008, pp. 368-372
Bioline Code: zr08058
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 29, No. 4, 2008, pp. 368-372

 en Construction and Analysis of A Directional Xenopus laevis check for this species in other resources Embryonic cDNA Library for Yeast Two-hybrid Using Modified Random Primers
WU, Jing-yang; LI, Chao-cui; KONG, Qing-hua & MAO, Bing-yu


Here we describe a new strategy for cDNA library construction, in which the random primers directing the first strand cDNA synthesis contain additional d (AC) at their 5'-ends, and the linkers which will be added to the double strand cDNA contain d(GTCG) at the 5′-ends. When the linker is added to the 3'-end of the cDNA, a complete SalI site d(GTCGAC) will form at the 3'-end but not at the 5′-end of the cDNA fragment. After SalI digestion, the 3'-cohesive and the pre-existing 5'EcoRI cohesive ends are used to introduce the double stranded cDNA into the linearized plasmid vectors. The ligation products were used to transform E.coli to produce a cDNA library. Using this method, we constructed a directional Xenopus laevis check for this species in other resources embryonic cDNA library for yeast two-hybrid. We checked the ratio of empty vectors, the size of inserts and existence of several genes, the results showed that cDNA library construction was successful.

Xenopus laevis; Yeast two-hybrid; Random primer; Directional cDNA library

© Copyright 2008 Kunming Institute of Zoology, the Chinese Academy of Sciences
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