RFLP subtraction

[Takashi Hashimoto]

I am interested in applying "RFLP subtraction" (Straus lab, PNAS 91:
6113-6117) on Arabidopsis genome.
1) Does anyone have experience with this method?  Does it work?  How is it
compared with representational difference analysis (RDA; Lisitsyn et al.,
Science 259: 946-951) in generating RFLPs?
2) In the original protocol of RFLP subtraction, one PCR primer is labeled
with biotin at two positions (one 5' end and one internal position).  Here
in Japan, I have been unable to find a commercial company that does
internal biotin-labelling service.  Is double labelling of the primer
necessary for efficient subtraction?
3) Maybe I can address these questions to Donald Straus of Brandeis
University himself.  Does anyone know his e-mail address?
Thank you for any advice.  Takashi
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Dr. Takashi Hashimoto
Graduate School of Biological Sciences
Nara Institute of Science and Technology
8916-5 Takayama-cho, Ikoma
Nara 630-01, Japan

Fax: 81-7437-2-5489
e-mail: hasimoto at bs.aist-nara.ac.jp
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