probs digesting plant genomic

QIAGEN qiagen at kaiwan.com
Wed Feb 7 20:05:00 EST 1996


BOTPG at LUFF.LATROBE.EDU.AU (GRANT,Patricia) wrote:

>I'm trying to digest genomic DNA from mature leaf tissue for eventual 
>ligation and use in inverse-PCR. I have been having problems trying to get 
>the stuff to digest with the restriction enzymes of choice (PvuII or XhoI). 
>I've tried several methods of genomic isolation including a quick and dirty 
>arabidopsis method (thought its not arab I'm working on), then CsCl gradient 
>then CTAB. None of these produced DNA which would cut with either of the 
>enzymes. Finally I have included Qiagen columns (anion exchange) as the final 
>purification step and it now cuts when I use BamHI but won't with either of 
>the others. Does anyone have any suggestions....? 

For more specific plant information from QIAGEN, please call our Technical 
Services Dept. at (800) 362-7737 from within the US or 02103-892-240
internationally to request our genomic plant protocol for genomic tips or
QIAamp.  If these procedures are not acceptable, another option is to do a
phenol/chloroform extraction and then use QIAGEN Genomic #10223. There is
a paper using this method.

K. Weising, B. Beyermann, J. Ramser, and G. Kahl (1991)                 
Plant DNA Fingerprinting with Radioactive and Digoxigenated Oligonucleotide 
Probes Complementary to Simple Repetitive DNA Sequences       
Electrophoresis 12, 159­169               
Genomic DNA isolation from plant (50 different species) on QIAGEN-tip 20.

-- 
QIAGEN, Inc.
1-800-426-8157
Temporary e-mail address: qiagen at Kaiwan.com



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