I have to prepare a percoll additive as follow: in a g/20ml
CaCl2.2H2O. ( 0.616 g)
MgCl2. ( 0.118 g)
NaCl. (0.935 g)
KCl. (0.046 g)
NaH2PO4. ( 0.009 g)
Gentamycin. (1ml)
Hepes Free Acid. ( 0.2384 g)
Hepes Na+ Salt. ( 0.2602 g)
Na Lactate Syrup ( 0.74 g)
I tried different ways to help not performing cloudiness.
1- add all the chemicals into 16 ml distilled water except for CaCl2.2H2O; all dissolved perfectly.
2- adding the CaCl2.2H2O into 1.5 distilled water; it dissolved perfectly.
3 - adding the 1.5 ml dissolved CaCl2.2H2O into the previous mixture, and unfortunately, it performing a cloudy mixture.
By the way, I tried to add CaCl2.2H2O and NaH2PO4 into distilled water, and it dissolved perfectly. Then once adding them into the other mixed chemicals, it performs the cloudy color. Then I tried the water bath sonication for about 1 and half hours; it dissolved a bit but not completely.
Can anyone give me the solution, please?
Other question:
Is there another gradient system rather than percoll that I can use for bovine sperm preparation?
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