Help with DNA isolation

Michelle Gleeson michelle at MOLECULE.BIO.UTS.EDU.AU
Sun Jul 7 16:42:23 EST 1996



On 5 Jul 1996 R.Peeters at nunhems.nl wrote:

> We have a problem with the preparation of high quality DNA from polyphenolic containing plants such as chicory. The DNA isolation method we use now is a
> standard CTAB isolation with 0,5% sodiumbisulfite and 1% PVP. A brown coloured substance often copurified with the chicory DNA even after rigorous purification.
> The brown material interferes with the PCR and gives bad, and sometimes none, bandingpatterns.
>
> Can anyone send us a fast DNA isolation method to avoid polyphenol contamination of the final DNA preparation.
>
> --------------------------------------
> Dr. R.A. Peeters
> Biotechnology department
> Nunhems Zaden BV
> POBOX 4005, 6080 AA, HAELEN
> Voort 6, 6083 AC, NUNHEM
> The Netherlands
> phone  : (+31) 475 599222
> fax    : (+31) 475 599223
> e-mail : R.Peeters at nunhems.nl
> --------------------------------------
>
> 
hi,
I used a method for extracting total nucleic acids from citrus which was a
combination of two papers:  Dellaporta, Wood,and Hicks (1983) Plant
Molecular Biology Reporter Vol 1. No 4 p 19-21 and Rowhani, CHay,
Golino and Falk (1993) Phytopathology Vol 83, No 7 p749-753 "Development
of a Reverse Transcriptase PCR Technique..."..  These methods gave me very
clean DNA, whereas  the previous method always gave me brown DNA which
inhibited the PCR.  This method takes about 1 day to do.  Good luck,
Michelle Gleeson
michelle.gleeson at uts.edu.au  > >




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