6 replies to this topic

[anonymous]

Sigma sells a trireagent specially for blood.

[anonymous]

Does anyone have a method for RNA extraction from whole blood and the expected yields?Has anyone used Trizol for RNA from blood?

[anonymous]

We just tried to extract RNA from whole blood with Trizol and unfortunately the result was decieving, we didn't get any RNA!

[romaoleao]

It is possible to preserve and do RNA extraction correctly in buffy coat after its fresh frozen in liquid nitrogen and cyopreservation at -80ºC? Is is necessary to add any preservation medium as RNA later or fresh frozen is just enough?

[findik44]

Hi evryone,

Just last week I tried the Tri-reagent to extract RNA from PBMCs containing virus in it and got no RNA. we usually use Trireagent for RNA extraction and  gives very high yiled. But this one was my first trial with PBMCs and I don`t know what went wrong?

[findik44]

anonymous said:

Sigma sells a trireagent specially for blood.

I would not suggest to do the RNA extraction from whole blood unless it is necessary. If so you can still use trireagent but as far as I know because of potential contamination risk you would have to do some sort of purification step afterwards.

[AquaPlasmid]

I can attest to Anonymous’s finding that total RNA could not be extracted from whole blood, especially frozen blood, collected in common anticoagulants, using Trizol. The standard method for blood RNA extraction is to lyse the RBC in fresh blood within 48 hrs with a hypotonic solution and then extract RNA from the enriched WBC with Trizol or other RNA kits. But such WBC enrichment step has its limitations. It could alter the expression profile of your target gene, and cause the loss of WBC, viral RNA, and cell-free circulating miRNA. Would it be nice if we could freeze the whole blood and then extract RNA from the frozen blood? But freezing introduces even more problems: the leaky RBC can no longer be removed, freezing damages the WBC and exposes the RNA to RNases, which is abundant in the RBC and the plasma, furthermore, free RNA binds to the hemoglobin and get lost during purification. The reality is that no method could actually recover total RNA from anticoagulated frozen whole blood. Now the good news: AquaPreserve http://www.aquaplasm...uaPreserve.html. For this method, you just mix the whole blood with AquaPreserve, precipitate the proteins with ProSink, and then precipitate total blood DNA and RNA from the clear lysate with isopropanol. We think we might have solved a big problem and allow researchers to use frozen whole blood samples obtained from routine testing, clinical trials, or epidemiological studies not only for DNA but also RNA studies.

Edited by AquaPlasmid, 04 June 2012 - 10:15 PM.