A kind of fetal bovine serum substitute and its preparation method and application
A technology of fetal bovine serum and substitutes, applied in biochemical equipment and methods, microorganisms, tissue culture, etc., can solve the problems of high production and use costs, and achieve good growth status and good inter-batch stability
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[0049] A kind of fetal calf serum substitute, its preparation method is as follows:
[0050] 1) Choose robust adult cows, take 5 liters of blood as a unit, add anticoagulant (4% trisodium citrate dihydrate) to 500ml; put it in a refrigerator at 4°C overnight, and take yellow plasma under sterile conditions , centrifuge at 3000rpm at 4°C for 10min and take the supernatant;
[0051] 2) Add CaCl with a final concentration of 20mM to the supernatant 2 , 37°C water bath for 2h;
[0052] 3) After standing at -20°C for 12 hours, place at room temperature to melt, remove the protein precipitate and collect the supernatant;
[0053] 4) Add NaCl with a final concentration of 4.79M to the supernatant, and precipitate at 4°C for 12 hours;
[0054] 5) Centrifuge at 5000rpm for 15min and collect the supernatant;
[0055] 6) Desalting, adjusting the NaCl concentration to 0.7%;
[0056] 7) Add ions necessary for cell growth, and the final concentration is:
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