HELP! muscle RNA extraction - (May/08/2006 )
Hi! I have a problem. I have to isolate RNA from a sample of muscle that weight 6 mg so I can´t used ultraturrax and disruption of myofibers using only a pellet pestle is veru difficult.
Can somebody help me, please?
well... do you use a kit? a tri reagent?...
I think that you may freeze your tissue and crush it / or add the lysis solution pre-heated up to 50°
Use the appropriately sized qiagen kit for the amount of starting tissue. Homogenise your sample using a rotor-stator/ polytron homogenizer. You need to follow their method for difficult tissues such as skeletal muscle which adds a proteinase K digestion before you attempt to add the homogenized sample on to the column. In the past I have isolated very good quality total RNA using this method.
Hope this helps
animal tissu is rich of ribonuclease use trizol reagent ( Invitrogen) or TRI reagent (SIGMA)