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Old 03-24-2014, 06:26 PM   #1
hweeing
Junior Member
 
Location: St Lucia

Join Date: Feb 2014
Posts: 4
Default PBMC extraction for RNA-seq

i'm very new to RNA-seq and I'm trying to extract PBMC for RNA sequencing.

I know that there is the ficoll technique to isolate PBMC using different density gradient centrifugation method, but this technique can is too long for me to extract sufficient RNA and the process might alter the RNA that might not be reflective of the condition.

Is there anyone out there that has another recommendation?
is it more suitable if i extract rna from whole blood since there is no DNA in red blood cells and they cannot synthesise rna.

Last edited by hweeing; 03-24-2014 at 06:57 PM.
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Old 03-26-2014, 11:31 AM   #2
JamieHeather
@jamimmunology
 
Location: London

Join Date: Nov 2012
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Default

There are things like PAXgene and Tempus tubes for extraction of DNA or RNA from whole blood, which just lyse everything before nucleic acid purification

That said, while RBC lack nuclei they are still jam-packed with globin mRNA; working from whole blood often requires depleting this to get any signal.
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