Method for simultaneously detecting genomic DNA polymorphism and methylation
A DNA polymorphism and genome technology, applied in the field of molecular biology, can solve the problems of time-consuming and laborious, unable to meet the direct connection and other problems
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[0026] Example 1 The method for simultaneously performing genomic DNA polymorphism and methylation detection
[0027] The technical route of the present invention is as figure 1 As shown, the specific method is:
[0028] 1) DNA fragmentation
[0029] Take 3 μg of genomic DNA from 97L liver cancer cells (MHCC-97L cells, purchased from Shanghai Bangjing Industrial Co., Ltd.), add water to adjust the volume of DNA to 85 μL, and use a Biorupter to disrupt DNA at 4°C; power: 80%, working time 30s , the gap time is 30s, 6 cycles, the DNA fragment is interrupted to about 300bp (such as figure 2 , where ultrasonic fragmentation of genomic DNA to a major band of 300 bp size can be seen).
[0030] 2) 3' end enzyme digestion treatment
[0031] 1. Prepare the terminal repair system by operating on ice, and add in the following order:
[0032] dna
35μL
10×NEB Buffer2 (purchased from NEB, item number catB7002)
10 μL
Klenow
5μL
wxya 2 o
...
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