Xin protocol tách chiết DNA plasmid từ Bacillus.

cafemuathu

New member
#1
Em chào cả nhà, hiện tại Em đang gặp khó khăn trong việc tách DNA plasmid từ Bacillus, Em sử dụng quy trình:
1. Sol I ( 25% sucrose, 50mM EDTA, 50 mM Tris-HCl pH 8.0) and 5mg of lysozyme per ml.
Sử dụng 200ul hào tế bào, sau đó ủ ở 37 độ 10'.
2. Sử dụng sol II, sol III, (25:24:1), (24:1) như tách chiết DNA plasmid từ E.coli.
Em sử dụng quy trình:

http://aem.asm.org/cgi/reprint/59/4/1138.pdf

Cả nhà giúp Em với ah!
 

Biologist

Member
Google "DNA extraction from Bacillus"

http://www.bio.net/bionet/mm/methods/2003-July/096531.html

To obtain genomic DNA, 1.5 ml of each overnight culture
was centrifuged at 13,000 g for 3 min. The pellets were resus-pended
in 1 ml of 10 mM Tris HCl, 10 mM EDTA, 100 mM
NaCl, 2% (v/v) SDS, and 400 mgml –1 proteinase K; the mix-ture
was incubated at 56°C for 30 min. After phenolization
and chloroform purification steps, DNA was precipitated
with 1 volume of 2-propanol, washed twice with 70% (v/v)
ethanol, and finally resuspended in 200 ml of sterile water.
 

cafemuathu

New member
Google "DNA extraction from Bacillus"

http://www.bio.net/bionet/mm/methods...ly/096531.html

To obtain genomic DNA, 1.5 ml of each overnight culture
was centrifuged at 13,000 g for 3 min. The pellets were resus-pended
in 1 ml of 10 mM Tris HCl, 10 mM EDTA, 100 mM
NaCl, 2% (v/v) SDS, and 400 mgml –1 proteinase K; the mix-ture
was incubated at 56°C for 30 min. After phenolization
and chloroform purification steps, DNA was precipitated
with 1 volume of 2-propanol, washed twice with 70% (v/v)
ethanol, and finally resuspended in 200 ml of sterile water.

Em cám ơn Bác David Dang nhé,
Để em thử phương pháp này xem thế nào, kết quả thế nào nhờ các Bác chỉ giáo tiếp cho em với ah!
 

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