Thaw
The serum (Fetal Bovine Serum, Hyclone) was taken out from the -20 ° C refrigerator, placed in a refrigerator at 4 ° C for overnight freezing; it can also be frozen at room temperature (or immersed in tap water), and if it is not immediately inactivated after defrosting, it can be placed in a refrigerator at 4 ° C. Temporarily saved;
2. Inactivation
(1) Start heating in a water bath at room temperature (put in a serum bottle containing 200 ml of tap water, insert a thermometer, temperature monitoring);
(2) When the thermometer shows 56 °C, control at this temperature for 30 minutes ± 3 minutes; if the temperature rises above 56 °C, if it does not exceed 60 °C, and the time is very short (for example: no more than 5 minutes) ), can still be used; if it exceeds 60 ° C, or for a long time, discard it, or try to culture the ME cells;
(2) Take out, naturally cool to room temperature (about 1-3 hours), can be divided or -20 °C to continue to freeze;
3. Packing
(1) Transfer to the cells, pipette the serum, pay attention to the serum should be gently shaken for several weeks, mix; note when blowing out the serum: do not blow out bubbles (the serum is very viscous, it is easy to produce bubbles If the bubble has been generated, pass it on the flame of the alcohol lamp); mark the lot number and date;
Shanghai Chuangsai Technology provides a lightweight P-type pipette (set one: 10ul, 100ul, 1000ul), product number: PNT01, price 5,000 yuan.
(2) Store in a refrigerator at -20 °C (approx. 1-2 months).
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