miniprep of plasmid DNA
- 최초 등록일
- 2014.09.13
- 최종 저작일
- 2014.08
- 3페이지/ 한컴오피스
- 가격 1,000원
목차
1. Principle & Key Steps
2. Materials
3. Protocol
4. References
본문내용
1. Principle & Key Steps
1) Alkaline lysis: isolation of plasmid DNA from E.coli ( Birnboim and Doly 1979) works well with bacterial cultures ranging in size from 1ml to >500ml.
2) Denature: Suspensions to the strongly anionic detergent at high pH opens cell wall, denatures chromosomal DNA and protein, and release plasmid DNA into the supernatant.
3) Neutralization: Bacterial proteins, broken cell walls, and denatured chromosomal DNA are efficiently precipitated from solution when sodium ions are replaced by potassium ions.
<중 략>
1) Binding: variable salt and pH
2) Elution: variable salt and pH
3) Alcohol precipitation
1) Binding: high salt
?Nucleic acids are adsorbed to the silica?gel membrane in the presence of chaotropic salts, which remove water from hydrated molecules in solution. Polysaccharides and proteins do not adsorb and are removed.
참고 자료
J. Sambrook and D. W. Russell., 2001., Molecular cloning, the third edition., CSHL press., Vol 1, pp 1.31‐1.34
QIAGEN., 2005., QIAGEN plasmid Purification Handbook., the second edition., pp 15‐18