- This topic has 4 replies, 2 voices, and was last updated April 5, 2020 at 8:36 pm by .
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Hi everyone,
this is the main discussion space for adjusting BOMB protocol #8.2 RNA extraction from mammalian cells using GITC lysis. for use in COVID-19 sample testing.
Older posts concerning this COVID-19 specific adjustment have been moved from the general RNA extraction Forum to this new topic.
This protocol seems very useful for my desire to develop an open source, do-it-yourself, protocol for extracting COVID-19 viral RNA from samples. Every time another commercial kit is approved, it is backordered for an unknown amount of time. I am a University professor who is shut out of his own lab that happens to know the CEO of a local biotech company. They are trying to ramp up the capability to become a testing lab for COVID-19 samples. We are in Oklahoma, and are about 1-3 weeks behind the curves on the East and West coasts. Given what shortages they are experiencing now, by the time we need to be able to test widely (weeks ago in my opinion), the resources to do so will be even more scarce. We are hoping that if we can obtain the raw materials for the extraction of viral RNA using GITC and magnetic beads, we could adapt this to multiple automated platforms. We are most interested in the Opentrons platforms, as we are 100% interested in sharing our successes with the worldwide community so that everyone can benefit. Let’s democratize testing for COVID-19 because we are all in this together as a species.
Ahoy,
I fully agree and support your endeavor.
We have already had two inquiries of the same/similar kind. Please have a look here for some more information:
BOMB protocol #8.1 RNA extraction from mammalian cells using TRI reagent
Stay Safe.
Tim M
Hi,
I am also working on COVID19 diagnostics, based in London. We are going to try to use the GITC lysis RNA extraction protocol on our samples because sourcing commercial kits is starting to become a serious issue. If we can make our own then hopefully that would be much better!
I wondered what volume of lysis buffer you would recommend adding to our samples? We cannot centrifuge to pellet the virus, so we have to start from a suspension of the virus in transport media (HBSS) the swabs come in. Or we could make a more concentrated version of the lysis buffer to dilute to 1x in the transport media? I know we need to keep the lysate:isopropanol:bead ratio consistent. Any help would be greatly appreciated.
Hi Rob,
we just released a protocol addressing exactly this!
https://bomb.bio/wp-content/uploads/2020/04/SARS-CoV-2-RNA-purification-from-nasal-swabs_BOMBv2.pdf
Cheers,
Phil
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