RNA extraction from skin samples

Geoff Scopes geoff at advbtech.demon.co.uk
Fri Dec 15 07:31:07 EST 1995


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 
>in touch.
>
>Thanks,  
>Ginny Shires
>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.

Best Regards
Russell Herbert
Advanced Biotechnologies Ltd
(01372) 360123
-- 
Russell Herbert

Turnpike evaluation. For information, email: info at turnpike.com



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