Interference fragment and application thereof
A fragment and gene technology, applied in the field of interference fragment and its application, can solve the problem that there is no cure for the root cause, achieve important market value and significance, and alleviate the effect of corresponding symptoms
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[0047] 1. Design the AQP1 siRNA sequence, and design the corresponding negative control siRNA, and then submit it to the gene company for synthesis; among them,
[0048] AQP1 RNAi sequence:
[0049] Positive sequence: 5'-GCUGUACUCAUCUACGACUTT-3';
[0050] Reverse sequence: 5'-AGUCGUAGAUGAGUACAGCTT-3'.
[0051] 2. Aliquot all AQP1 siRNA sequences and negative control interference sequences into 1OD / tube, centrifuge at 3000-4000rpm for 3min before use; then add 125μL DEPC H 2 O resuspended to prepare a stock solution with a final concentration of 20 μM.
[0052] 3. Human umbilical vein endothelial cells (HUVEC cells) use improved complete medium (MCDB131 medium + 20% fetal bovine serum + 50mg / mL endothelial growth factor ECGS + 100U / mL penicillin + 50μg / mL heparin + 2mmol glutamine +100U / mL Streptomycin) after recovery, trypsinized and replanted in 6-cell culture plate; 37°C, 5% CO 2 After culturing overnight, start the cell transfection experiment when the coverage rate of th...
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