Method for analyzing an integration site of lentiviral vector in CAR-T cells, and primers
A technology of lentiviral vector and analysis method, which is applied to the analysis of the integration site of lentiviral vector in CAR-T cells. The primer sequence field of this method achieves the effect of good specificity, stability and high sensitivity.
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 Example 1 Analysis of integration sites of lentiviral vectors in CAR-T cells
 The primer sequences are shown in the table below:
 1 Experimental method
 1.1 Extraction of total genome DNA of anti-BCMACAR-T cells
 (1) Sample homogenate
 Add 1ml DNAZOL Reagent to the CAR-T cell sample, and pipette 50 times, taking care to avoid air bubbles.
 (2) DNA precipitation
 Add 0.5ml of 100% ethanol to the above cell homogenate, turn it upside down 10 times, and let it stand at room temperature for 1-3 minutes, flocculents can be seen. Centrifuge at 15000g for 5min. Remove the supernatant with a pipette.
 (3) DNA washing
 Add 1ml of 75% ethanol, invert up and down 10 times, and centrifuge at 12000g for 1min. Carefully remove the supernatant with a pipette. repeat. Open the cap of the EP tube for 3-5 minutes, and let it dry in the air.
 (4) DNA dissolution
 According to th...
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