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2010•Journal of Peanut ScienceRequires access

Construction of a Full-length cDNA Library from Peanut Seedlings

Zhen Yang

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Abstract

In order to study the novel genes of peanut, a cDNA library from seedlings was constructed. Total RNA was extracted from E12 peanut seedlings and mRNA was purified. Double strand cDNA was synthesized by SMART method, the ds cDNA fragments were ligated into the pBluescriptⅡSK vector. The recombinant plasmids were transformed into the E. coli, a cDNA library of peanut seedlings was successfully constructed. The titer of the cDNA library was estimated as 1.1×106cfu/mL; the percentage of recombination was 93.4%. PCR results showed that the inserts varied from 0.75 to 2.0 kb.

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What this paper is about

In order to study the novel genes of peanut, a cDNA library from seedlings was constructed. Total RNA was extracted from E12 peanut seedlings and mRNA was purified. Double strand cDNA was synthesized by SMART method, the ds cDNA fragments were ligated into the pBluescriptⅡSK vector. The recombinant plasmids were transformed into the E. coli, a cDNA library of peanut seedlings was successfully constructed. The titer of the cDNA library was estimated as 1.1×106cfu/mL; the percentage of recombination was 93.4%. PCR results showed that the inserts varied from 0.75 to 2.0 kb.

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Available abstract

In order to study the novel genes of peanut, a cDNA library from seedlings was constructed. Total RNA was extracted from E12 peanut seedlings and mRNA was purified. Double strand cDNA was synthesized by SMART method, the ds cDNA fragments were ligated into the pBluescriptⅡSK vector. The recombinant plasmids were transformed into the E. coli, a cDNA library of peanut seedlings was successfully constructed. The titer of the cDNA library was estimated as 1.1×106cfu/mL; the percentage of recombination was 93.4%. PCR results showed that the inserts varied from 0.75 to 2.0 kb.

Key concepts: Complementary DNA, cDNA library, Recombinant DNA, Biology, Titer, Plasmid, Molecular biology, RNA

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