| Literature DB >> 12885174 |
Yoon Kwon Nam1, Ji Eun Park, Kyoung Kil Kim, Dong Soo Kim.
Abstract
The protocol described in this paper offers a simple and rapid method for PCR analysis of transgenes using a restricted amount of fin tissue from small-sized transgenic fish. A simple preparation of fin lysate using a buffer containing a low concentration of an ionic detergent, SDS (0.01%), followed by neutralization with a second buffer containing higher concentrations of non-ionic detergents NP40 (2%) and Tween 20 (2%) consistently provides a reliable quantity of high-quality DNA template for PCR amplification of transgenes. Based on this protocol, transgenic fish can be clearly distinguished from non-transgenic fish using PCR in a rapid and reproducible manner. Tedious DNA purifications are avoided while fidelity of amplification and efficient identification of transgenic fish are maintained.Entities:
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Year: 2003 PMID: 12885174 DOI: 10.1023/a:1024274508052
Source DB: PubMed Journal: Transgenic Res ISSN: 0962-8819 Impact factor: 2.788