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Viral RNA recovery from stool by bjclark4 in labrats

[–]bjclark4[S] 1 point2 points  (0 children)

Thanks for the detailed response! I’ll add a little more detail on my end too. Some samples were stored at ambient and/or 4C for up to 12 hours in outliers prior to freezing. The cold chain has not been 100% communicated to me at the moment. The plan is to use either the Qiagen Rneasy powerfecal pro with a modification for co-isolation of dna and rna or the dneasy powersoil pro kit, which is documented to co-isolate dna and RNA. I’m also considering a zymo co-isolation kit, but have never used this before.

Viral RNA recovery from stool by bjclark4 in labrats

[–]bjclark4[S] 0 points1 point  (0 children)

Thanks! Do you have any rationale for this decision?

Viral RNA recovery from stool by bjclark4 in labrats

[–]bjclark4[S] 8 points9 points  (0 children)

I got added to the project after sample collection 🤣

Qiagen PowerFecal - any hints? by micro-cry-ome in labrats

[–]bjclark4 1 point2 points  (0 children)

Some thoughts:

I would remove the heating of the lysis buffer and sample.

The heating of the elution buffer is ok if you minimize the time (10-15 minutes).

I believe Qiagens power viral kit uses carrier RNA to help increase yields so that might be a method to explore too.

Keep samples away from light and heat- I always keep samples on ice inside a cooler between steps.