title: A rapid and simple PCR-based method for analysis of transgenic fish using a restricted amount of fin tissue.
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.
authors.affiliation: Fish Genetic Manipulation Laboratory CNR), Department of Aquaculture, Pukyong National University, Pusan 608-737, South Korea. yoonknam@pknu.ac.kr
authors.forename: Yoon Kwon
authors.initials: YK
authors.lastname: Nam
authors.forename: Ji Eun
authors.initials: JE
authors.lastname: Park
authors.forename: Kyoung Kil
authors.initials: KK
authors.lastname: Kim
authors.forename: Dong Soo
authors.initials: DS
authors.lastname: Kim
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers; D005399:Fishes / Q000235:genetics*; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers