Construction and Identification of a Full-length cDNA Library from Peanut Seeds
Zhuang Wei-jian
Abstract
Zhuang Wei-jian
Abstract
In order to study the novel genes of peanut seeds,a full-length cDNA library from peanut seeds was constructed.mRNA from peanut seeds of different developmental stages was purified.Double strand cDNA was synthesized by SMART method,the ds cDNA fragments were ligated to the pDNR-LIB vector.The recombinant plasmids were transformed into the E.coli,a cDNA library of peanut seeds was successfully constructed.The full-length cDNA library was stocked after amplification,the titer of the cDNA library was estimated as 1.7×109 cfu/mL.PCR results showed that the inserts varied from 0.5 to 2.0kb with average size was larger than 1000bp.It indicated that this library could be used for full-length genes screening and cloning of low abundance genes.
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In order to study the novel genes of peanut seeds,a full-length cDNA library from peanut seeds was constructed.mRNA from peanut seeds of different developmental stages was purified.Double strand cDNA was synthesized by SMART method,the ds cDNA fragments were ligated to the pDNR-LIB vector.The recombinant plasmids were transformed into the E.coli,a cDNA library of peanut seeds was successfully constructed.The full-length cDNA library was stocked after amplification,the titer of the cDNA library was estimated as 1.7×109 cfu/mL.PCR results showed that the inserts varied from 0.5 to 2.0kb with average size was larger than 1000bp.It indicated that this library could be used for full-length genes screening and cloning of low abundance genes.
Key concepts: Complementary DNA, cDNA library, Biology, Plasmid, Rapid amplification of cDNA ends, Recombinant DNA, Cloning (programming), Molecular biology