Hi,
I usually use sucrose lysis buffer ( 0.6 M sucrose, 0.02 M Tris base, 0.01 M MgCl2 and 2% triton) for RBCs lysis in DNA extraction. After initial RBCs lysis and washing, a pellet of WBCs is obtained upon which extraction is completed.
Could I use this buffer for isolation of WBCs as a source of RNA or protein as well???
what could be the operating and centrifugation conditions?
Hello, I think that sucrose was used more extensively for DNA isolation probably because this material is more stable than RNA. Anyway since sucrose is a suitable tool for subcellular fractionation and for retention of isotonic osmolarity I guess it could work for RNA isolation too. Also, to optimize your extraction I think you have to add a nuclease to digest DNA since you don't need it and a protease to digest proteins and finally an RNase inhibitor to protect your RNA especially considering the quick degradation of RNA.
Hello, I think that sucrose was used more extensively for DNA isolation probably because this material is more stable than RNA. Anyway since sucrose is a suitable tool for subcellular fractionation and for retention of isotonic osmolarity I guess it could work for RNA isolation too. Also, to optimize your extraction I think you have to add a nuclease to digest DNA since you don't need it and a protease to digest proteins and finally an RNase inhibitor to protect your RNA especially considering the quick degradation of RNA.
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