Full-Length Normalization Subtractive Hybridization: A Novel Method for Generating Differentially Expressed cDNAs |
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| Authors: | Zhong-Min Dai Xiao-Jing Zhu Wei-Jun Yang |
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| Institution: | (1) Institute of Cell Biology and Genetics, College of Life Sciences, Zhejiang University, Zijingang Campus, Hangzhou, Zhejiang, 310058, People’s Republic of China;(2) State Conservation Center for Gene Resources of Wildlife, Key Laboratory of Conservation Genetics and Reproductive Biology for Wild Animals of the Ministry of Education, Hangzhou, Zhejiang, 310058, People’s Republic of China; |
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| Abstract: | Here we present a novel method termed full-length normalization subtractive hybridization (FNSH) for efficiently generating
subtracted cDNA libraries with a high degree of productivity. This method has the ability to isolate full-length differentially
expressed genes from target samples. Normalization and subtraction of FNSH are performed simultaneously with efficiency equal
to or even higher than that of suppression subtractive hybridization. Using FNSH, we have isolated at least 40 unique cDNAs
that are expressed in terminal ampullae but not in the ovaries of the prawn Macrobrachium rosenbergii from 120 randomly picked subtracted clones. Sequence analysis shows that 37 of the 40 cDNAs are full length. |
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