Literature DB >> 12885174

A rapid and simple PCR-based method for analysis of transgenic fish using a restricted amount of fin tissue.

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.

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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


  3 in total

1.  Isogenic transgenic homozygous fish induced by artificial parthenogenesis.

Authors:  Y K Nam; Y S Cho; D S Kim
Journal:  Transgenic Res       Date:  2000-12       Impact factor: 2.788

2.  Dramatically accelerated growth and extraordinary gigantism of transgenic mud loach Misgurnus mizolepis.

Authors:  Y K Nam; J K Noh; Y S Cho; H J Cho; K N Cho; C G Kim; D S Kim
Journal:  Transgenic Res       Date:  2001-08       Impact factor: 2.788

3.  Identification of transgenic mice by direct PCR analysis of lysates of epithelial cells obtained from the inner surface of the rectum.

Authors:  H Lahm; A Hoeflich; N Rieger; R Wanke; E Wolf
Journal:  Transgenic Res       Date:  1998-03       Impact factor: 2.788

  3 in total
  3 in total

1.  Characterization of stable fluorescent transgenic marine medaka (Oryzias dancena) lines carrying red fluorescent protein gene driven by myosin light chain 2 promoter.

Authors:  Young Sun Cho; Sang Yoon Lee; Dong Soo Kim; Yoon Kwon Nam
Journal:  Transgenic Res       Date:  2012-11-28       Impact factor: 2.788

2.  Characterization of estrogen-responsive transgenic marine medaka Oryzias dancena germlines harboring red fluorescent protein gene under the control by endogenous choriogenin H promoter.

Authors:  Young Sun Cho; Dong Soo Kim; Yoon Kwon Nam
Journal:  Transgenic Res       Date:  2012-09-13       Impact factor: 2.788

3.  Functional ability of cytoskeletal β-actin regulator to drive constitutive and ubiquitous expression of a fluorescent reporter throughout the life cycle of transgenic marine medaka Oryzias dancena.

Authors:  Young Sun Cho; Sang Yoon Lee; Youn Kyoung Kim; Dong Soo Kim; Yoon Kwon Nam
Journal:  Transgenic Res       Date:  2011-03-25       Impact factor: 2.788

  3 in total

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