[유전학] Mini prep
- 최초 등록일
- 2015.12.28
- 최종 저작일
- 2014.12
- 5페이지/ MS 워드
- 가격 2,000원
목차
1. Method
2. Result
3. Discussion
Reference
본문내용
* Method
Maxiprep-Alkaline Lysis(100ml)
- Preparation of cell culture
1. Pick colony and grow in 3ml LB over night at 37℃
2. Inoculate 100ml LB with antibiotics and grow over night at 37℃
3. Centrifugation at 2700G (4100rpm in a Sorvall GSA rotor) for 15min at 4℃
4. Resuspend pellet in 40ml cold STE buffer
5. Centrifugation as described in step 3
- Lysis of cells
6. Resuspend pellet in 4ml ALS-I buffer
7. Add 400μl Lysozym(10mg/ml)
Mix well by swirling the tube
8. Incubate for 5 min at room temperature
9. Add 8ml ALS-II
Mix thoroughly by inverting the bottle several times
10. Incubate for 10 min at room temperature
11. Add 6ml ice cold ALS-III buffer
Mix the contents gently
12. Incubate for 10 min on ice and mix again by swirling
13. Centrifuge at ≥20,000g (11,000rpm in a Sorvall GSA rotor) for 30 min at 4℃
- Recovery of plasmid DNA
14. Filter the supernatant through 4 layers of cheesecloth/miracloth
15. Add 0.6volume of Isopropanol
참고 자료
분자 생물학 노트, KIST생명공학연구소, 한림워느 1998,
유전공학개론. Takeshi Uozumi, 청문각, 2001,
생물공학 실험서, 주정희, 자유아카데미, 2000,
분자생물학실험서, 김영희, 월드사이언스, 2011,
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