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HELP HELP HELP needed with RNA extraction

Dimple Win ez059690 at peseta.ucdavis.edu
Tue May 21 21:47:29 EST 1996


On 13 May 1996, Eric Sarnighausen wrote:

> I am currently trying (again and again.....) to extract RNA from woody samples
> (Cornus sericea L.). I sterilize all the my equipment, use autoclaved and DEPC-
> treated water, buffers and solutions and wear gloves during the extraction
> procedure. The extraction buffer contains 4 M guanidine thiocyanate and I use a
> cesium chloride gradient followed by phenol/chloroform extraction and ethanol
> precipitation (as described by Clausen and Apel [1991]. Plant Mol. Biol. 17(4):
> 669-678) for the isolation of RNA from the xylem of poplar trees).
> The purity of the RNA (E260/E280) is 2.0, but no matter whether I use fresh or
> freeze dried material: the RNA is always completely degraded.
> Any help on this matter would be greatly appreciated.
> 
>                        Thanks in advance
> 
>                                             Eric
> 
> 
 	How many times have you repeated/checked for degradation of RNA? 
Did you use EDTA in your loading buffer before running electrophoresis to 
check if RNA is degraded? Let me know if this helps.

Dimple
dawin at ucdavis.edu



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