Please can someone help me with RBC lysis buffer for red blood cells?? I have prepared one and used it but it didn't work ... The problem is that I currently dont have small filter needed for sterilization of the buffer so I auto-calved it . I don't know if that could damage something I ajusted th pH after autoclave, but when I used the buffer with the blood it couldn't break the RBC and I wasn't able to preform RNA extraction. Here is the protocol that I sued to prepare my RBC lysis buffer:
RBC lysis buffer 10x
1. Prepare 8,26 g of NH4Cl
2. Add 1,19 g of NaHCO3
3. Add 200 µL of EDTA [0,5 M, pH8]
4. Add a.d until 100 mL 5. Adjust pH to 7.3
6. Filter sterile
I would appreciate any kind of help!