Trying to find an effect alternative lysis buffer not that different to GITC that can ensure effective lysis of infected macrophages with Mycobacterium tuberculosis. If planning to extract RNA from intracellular bacteria lysate. The reason why GITC is used is because it helps to prevent activity of RNase enzymes not to damage the extract, but I worry that the lysis with GITC doesn’t seem to be effective enough and end up less intracellular bacteria material to extract RNA from.
We lyse the macrophage with 0.07% SDS and then neutralize the SDS with 6% BSA. It works well and M. tuberculosis grows in good condition. I think you proceed with 0.1% triton or you may also use SDS.Both works fine.
Hope this helps!
Good luck.
Thank you everyone for your informative comments and suggestions. All of them were very helpful!! and assisted me in choosing the correct raw macrophages lysis buffer.
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