Plasmid DNA purification (NucleoBond Xtra Midi) kit Macherey-nagel
1. cultivate and harvest bacterial cells
4500-6000xg 4℃, 15min
2. cell lysis
8mL buffer RES(4℃) vertex로 cells을 풀어준다
8ml buffer LYS 5번 이내로 inverting 많이하게 되면
RT, 5min 기다리는 동안 3번 진행
3. Equilibaration of the column and filter
12ml buffer EQU cells을 키운 삼각플라스크에 준비
4. Neutralization
8ml buffer NEU
mix by gently inverting 10-15 times 청색에서 흰색으로 변함
5. clarification and loading of the lysate
invert the tube 3 times
load lysate on Nucleobond Xtra column filter
6. washing
5ml Buffer EQU
7. washing
8ml buffer WASH filter를 제거하고 함
8. elution
5ml buffer ELU 새 tube 준비
9. Precipitation
3.5ml isopropanol
5-15000 x g 4℃, 30min
10. wash and dry DNA pellet
2ml 70% ethanol 넣은 후 1ml 버리고 1ml은 e-tube로 옮김
5-15000 x g RT, 5min
5-10min
11. Reconstitute DNA
Appropriate volume of TE buffer
200ul에 녹임
https://www.mybiosource.com/learn/testing-procedures/plasmid-isolation/
Plasmid Isolation - MyBioSource Learning Center
Introduction The term ‘plasmid’ was coined by Joshua Lederberg in 1952. Originally evolved from bacteria, plasmids are extrachromosomal genetic elements present in most species of Archae, Eukarya and Eubacteria that can replicate independently. Plasmid
www.mybiosource.com
https://www.labex.kr/goods/goods_view.php?goodsNo=1000000109
NucleoBond Xtra Midi
Labex(라벡스)
www.labex.kr
