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This experiment introduced the operation steps of concentrating and purifying lentivirus with PEG-8000 concentration method.
1. NaCl
2. PEG8000
1. High pressure sterilizer
2. 0.45 μM filter head
3. High speed centrifuge
4. Low temperature refrigerator
1. Prepare 5 X PEG8000 NaCl and weigh 8.766 g NaCl; 50g PEG8000 are dissolved in 200 ml Milli-Q pure water; Sterilize with autoclave at 121 ℃ for 30min; Store at 4 ℃ after sterilization.
2. 0.45 μ M filter head was used to filter lentivirus supernatant.
3. Add 7.5 ml of 5 X PEG8000 NaCl mother solution every 30 ml of filtered initial virus solution.
4. Mix every 20~30min for 3-5 times in total.
5. Place it at 4 ℃ overnight.
6. Centrifuge for 20min with high speed centrifuge at 4 ℃, 4000 g.
7. Suck and discard the supernatant, leave the tube standing for 1~2 minutes, and absorb the residual liquid.
8. Add an appropriate amount of lentivirus lysate to dissolve lentivirus precipitation.
9. The collected virus suspension is divided into 50 μl each and shall be stored in the finished product tube. Store at -80 ℃ after quick freezing with crushed dry ice.
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