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Full-Length Normalization Subtractive Hybridization: A Novel Method for Generating Differentially Expressed cDNAs

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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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Acknowledgment

We thank Chris Wood of the College of Life Sciences, Zhejiang University, for critical reading of this manuscript. This work was supported by the National Natural Sciences Foundation of China (40730212) and the “863” program of China (2007AA09Z426).

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Authors and Affiliations

  1. Institute of Cell Biology and Genetics, College of Life Sciences, Zhejiang University, Zijingang Campus, Hangzhou, Zhejiang, 310058, People’s Republic of China

    Zhong-Min Dai, Xiao-Jing Zhu & Wei-Jun Yang

  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

    Wei-Jun Yang

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  1. Zhong-Min Dai
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  2. Xiao-Jing Zhu
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  3. Wei-Jun Yang
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Correspondence to Wei-Jun Yang.

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Dai, ZM., Zhu, XJ. & Yang, WJ. Full-Length Normalization Subtractive Hybridization: A Novel Method for Generating Differentially Expressed cDNAs. Mol Biotechnol 43, 257–263 (2009). https://doi.org/10.1007/s12033-009-9198-0

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  • DOI: https://doi.org/10.1007/s12033-009-9198-0

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