In article <Pine.SGI.3.91.951214161119.14840A-100000 at tower.york.ac.uk>,
VL Shires <vls100 at york.ac.uk> writes
>>I have just started a PhD and have been trying to extract total RNA from
>skin samples from mice for RT-PCR analysis of cytokines. I know that
>other people have managed to do this type of analysis before as there is
>published data. My RT-PCR step is fine as my positive controls are
>working beautifully but the message from the skin samples is at best very
>weak. So if any one has any suggestions re: more efficient extraction
>methods (currently using Guanidium, CsCl gradient and phenol method) or any
>other helpful tips for this work I would be very grateful if they could get
>University of York
It could be possible to increase your yield of RNA by using Protienase K
and guanidine, or Fibrolise which is a blend of enzymes that effectivy
distupts animal tissue for the extraction of nucleic acids.
Then use a standard total RNA isolation reagent.
Samples can be made available at request.
Advanced Biotechnologies Ltd
Turnpike evaluation. For information, email: info at turnpike.com