Re: [求救]關於DNA fragmentation
以下是我在網路上找到的方法
我有用過是可以的 有興趣沒錢買kit的可以試試看
4x10(6次方) target cells collect the cell sample in 1.5 ml eppendorf tube
--> spin down
--> resuspend with 0.5 ml PBS in 1.5 ml eppendorf tubes 55ul of lysis buffer
(40 ml of 0.5 M EDTA / 5 ml of 1 M Tris-Cl buffer pH 8.0 /
5 ml of 100% Triton X-100 / 50 ml of H2O)
--> 4℃ 20min
--> 4℃ 12,000g 30 min
--> extract the supernatant with 1:1 mixture of phenol:chloroform
--> gentle agitation for 5 min
--> 4℃ 12,000g 5min
--> 2V cold Ethanol+ 1/10V NaoAc
--> 4℃ 12,000g 5min
--> 30ul of deionized water-RNase solution
--> 30 minutes at 37℃
--> Run the 1.2% gel at 5V for 5min before increasing to 100V
來源
http://hedricklab.ucsd.edu/Protocol/DNAFRAG.html
(Hedrick Lab, UCSD Cancer Center)
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