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KMID : 1094720100150050817
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Biotechnology and Bioprocess Engineering
2010 Volume.15 No. 5 p.817 ~ p.821
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A simple, rapid, efficient and inexpensive strategy for sequencing clones from cDNA libraries
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Nguyen Dinh Truong
Oh Youn-Shin
Dirisala Vijaya R.
Choi Ho-Jun
Park Keun-Kyu
Kim Jin-Hoi
Park Chan-Kyu
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Abstract
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In this report, we describe a simple, rapid, efficient and inexpensive strategy for sequencing inserted DNAs from clones of cDNA or gDNA libraries. This strategy uses PCR products directly amplified from transformed bacterial colonies, with universal primers within the vector. The method can be applied for sequencing cDNA or gDNA libraries with up to 4 ∼ 5 kb insert sizes, without overnight liquid culture or plasmid DNA preparation steps. We successfully used this method to analyze clones from full-length, enriched cDNA libraries. Although simple, following this strategy will significantly help researchers to avoid unnecessary steps in the analysis of a cDNA library.
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KEYWORD
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cDNA library, screening, sequencing, colony PCR
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