Southern Hybridization

목차

Ⅰ. Introduction
Ⅱ. Materials
Ⅲ. Procedure
Ⅳ. References

본문내용

Ⅰ. Introduction
The principle of hybridization analysis is that a single-stranded DNA or RNA molecule of defined sequence (the "probe") can base-pair to a second DNA or RNA molecule that contains a complementary sequence (the "target"), with the stability of the hybrid depending on the extent of base pairing that occurs. Experimentally, the analysis is usually carried out with a probe that has been labeled and target DNA that has been immobilized on a membrane support.
The first stage in a hybridization experiment is to immobilize the denatured nucleic acids on a suitable solid support. The labeled probe is then applied in a solution that promotes hybridization. After a suitable incubation, the membrane is washed so that any nonspecifically bound probe is removed, leaving only probe that is base-paired to the target DNA. By controlling the stringency of the washing conditions, decisions can be made about whether to target sequences that are 100% complementary to the probe, or allow some mismatching so that sequences with lower degrees of similarity are also detected. The latter approach (heterologous probing) is used to study related sequences in a single or more than one genome.

참고 자료

Frederick M. Ausubel 외, Current Protocols in Molecular Biology, John Wiley & Sons Inc., 2004, 2.10.1~2.10.3
Sambrook and Russel, Molecular Cloning 3rd edition vol. 1, 2001, 6.50~6.55