DNA from Herbarium specimens

BRENDA WINGFIELD - PERSONEEL BRENDA at WWG3.UOVS.AC.ZA
Tue Jul 11 05:56:58 EST 1995


We have done quite a few PCRs from herbarium material.  In my 
experience some of the best sucesses I have had is directly from the 
old material itself, that is do not extract the DNA just add the 
"mycelium".  We have reported on this a couple of times and it really 
works well.  My suggestion over and above not extracting is to use a 
smaller region.  That is rather than ITS1 to ITS4 (which I presume is 
what you are using) use ITS1 to ITS2 and then ITS3 to ITS4.  Another 
problem we have encountered which we have not been able to resolve is 
that ITS1 and ITS4 simply do not work with some ascomycetes that we 
are interested in.   You do not say what group of organisms you are 
working with.

Good luck

Brenda Wingfield
.
> To:            mycology at net.bio.net
> From:          lorelei at TELEPORT.COM (Lorelei Norvell)
> Subject:       DNA from herbarium specimens
> Date:          9 Jul 1995 15:25:44 -0700
> 
> I have been successful in extracting and amplifying nDNA (ITS1 + 5.8srRNA 
> + ITS2) from over 100 different dried herbarium specimens.  However, I 
> have found that the older the DNA, the greater problems I have 
> encountered.  Unfortunately it is imperative I extract DNA from type 
> specimens collected from 1910 to 1970.  While I have had some success 
> with the 1970 and 1956 collections, I am encountering problems with 
> degraded DNA.
> 	I would welcome advice with respect to
> [1]  Whether drying the specimens again before extraction would increase 
> success or further degrade the DNA (drying with no heat).
> [2]  Another procedure besides the hot CTAB procedure I have used
> [3]  Any other preparation advice.
> 
> Thanks.
> Lorelei Norvell
> University of Washington
> Department of Botany
> 
> 




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