Random Priming for Radiolabeling of DNA fragments
- 최초 등록일
- 2014.09.13
- 최종 저작일
- 2014.08
- 3페이지/ 한컴오피스
- 가격 1,000원
목차
A. Objectives
B. Components of random priming reactions
C. Materials
D. Method
E. Reference
본문내용
A. Objectives
To produce radioactively labeled DNA strands for the detection of target DNA or RNA sequences in various applications including Southern and northern blot hybridization.
B. Components of random priming reactions
1. DNA Polymerase
Use the Klenow fragment of E. coli DNA pol I, the klenow fragment lacks 5'->3' exonuclease activity, so that the radioactive product is synthesized exclusively by primer extension rather than by nick translation and is not subject to exonuclease degradation.
2. Radiolabel
Random priming reactions typically contain one radiolabeled [a-32P]dNTP (sp. act. 3000 Ci/mmole)and three unlabeled dNTP as precursor.
3. Primers
The length of the primers is crucial.
참고 자료
Sambrook and Russell. 2001. Molecular cloning third edition. Vol 2 : 9.7-8