Is it possible to isolate intact RNA in peripheral blood? - (Apr/29/2008 )

Dear all,

I would like ask if we can isolate the intact RNA from peripheral blood samples?
My friend used Trizol to extract RNA from whole. However, the intensity of 28S:18S always falls into 1:1.
Do anyone have this experience?

Thank you so much.

siuchi

When you isolate RNA from blood, you need to remove the red blood cells and/or purify the white blood cells first.

Three ways to do this:

a) Red blood cell lysis ... there are many solutions you can buy or make, that lyse the RBCs. Mix in equal ratio with the blood, sit for 5-15 min at RT then collect the WBCs by centrifugation. Add 5-10 ml lysis solution and repeat incubation and spin until no redness in the pellet. Then use Trizol on the WBC pellet.

Purify the WBCs using a gradient mix like Ficoll-Paque.

c) Collect blood into PAXGene tubes (Qiagen) and use the Qiagen PAXGene Blood RNA kit.

If you do not remove the RBCs, you will have many problems downstream - like with PCR etc.

Hope this helps,

AussieUSA.

When you isolate RNA from blood, you need to remove the red blood cells and/or purify the white blood cells first.

Three ways to do this:

a) Red blood cell lysis ... there are many solutions you can buy or make, that lyse the RBCs. Mix in equal ratio with the blood, sit for 5-15 min at RT then collect the WBCs by centrifugation. Add 5-10 ml lysis solution and repeat incubation and spin until no redness in the pellet. Then use Trizol on the WBC pellet.

Purify the WBCs using a gradient mix like Ficoll-Paque.

c) Collect blood into PAXGene tubes (Qiagen) and use the Qiagen PAXGene Blood RNA kit.

If you do not remove the RBCs, you will have many problems downstream - like with PCR etc.

Hope this helps,

AussieUSA.