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Preparation of a genomic library using a TA vector
Y Kawata1, S Yano, A K Thankappan
1Osaka National Research Institute, Agency of Industrial Science and Technology, Ikeda, Japan.
Preparative Biochemistry & Biotechnology
|March 9, 1999
Abstract:
An efficient and simple method for constructing a genomic DNA library is presented using a TA cloning vector. It is based on sonication cleavage of genomic DNA, blunting of the fragment ends with mung bean nuclease, and addition of a single 3'-deoxyadenylate with Taq DNA polymerase, followed by ligation with a TA vector. This method is useful for improving the quality of genomic libraries for organisms whose genomic DNA is not well digested with restriction enzymes owing to the presence of polysaccharides and/or DNA methylation.