Method for inserting site in clone provirus
A technology of insertion site and provirus, applied in the field of life science and biology, can solve the problems of cost, limited quantity and difficulty, and achieve high efficiency.
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[0029] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.
[0030] A method of cloning a proviral insertion site comprising the steps of:
[0031] Step 1) culture cell line, described cell line is BXH-2 mouse leukemia cell line, cultivates in conventional ASM medium and contains 5% CO 2 Incubate at 37°C in a humid environment;
[0032] Step 2) Genomic DNA was extracted, using the Tiangen Genome Extraction Kit to extract genomic DNA from the collected cells according to the operating instructions, and then the DNA dissolved in ultrapure water was stored at -30°C;
[0033] Step 3) carry out the primer extension reaction (APE-PCR) with adenosine as the end, and the primer extension reaction with adenosine as the end comprises the following steps:
[0034] 3.1) Enzyme digestion, use restriction enzyme mixture Hind III, Pvu II and Xho I (Thermo) to digest different genomic DNA, incubate at 37°C ...
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