Fat removing - (Dec/09/2005 )

Hi all,
I have problems with fat in my samples. I have found out that it inhibit my RT step. I usually isolate RNA by TriReagent and I can not distinquish fat layer at beginning. Have you some tips to remove fat during procedure? Or can you recommend me some kit for fatty samples.

Thanks

Fat is ugly and evil. It is yellow and gross.

It sits on top of your aqueous layer. You can try to chloroform extract it.

tat troubleshoot may help :

OPTIONAL: An additional isolation step may be required for samples with high content
of proteins, fat, polysaccharides or extracellular material such as muscles, fat tissue, and
tuberous parts of plants. Following homogenization, remove insoluble material from the
homogenate by centrifugation at 12,000 × g for 10 minutes at 2 to 8°C. The resulting pellet
contains extracellular membranes, polysaccharides, and high molecular weight DNA,
while the supernatant contains RNA. In samples from fat tissue, an excess of fat collects as
a top layer which should be removed. In each case, transfer the cleared homogenate
solution to a fresh tube and proceed with chloroform addition and phase separation as
described.