Separating polymorphic amplified fragments of the same size
bertheau at inapv.inapg.inra.fr
Sat Nov 27 10:16:59 EST 1993
We are currently ampifying 1 to 3 fragments (depending on the bacterial
strain we are studying) of 420 bp which appear polymorphic after restriction
(the summation of the size corresponds to 1-3 fragments).
We would like to sequence these fragments from several strains in order to
develop ASO probes. The possibilities seem to be:
- cloning the fragments and trying to find the polymorphic fragments but it
seems very fastidious...
- to separate the fragments and then to sequence them but how to do ?
* TGGE or DGGE but developing TGGE protocols may be time consuming
and I would like to avoid Etidium bromide staining for the effects
of UV on DNA
* chromatography ? ion exchange or size exclusion ? Which material
is then available for such a work e.g. for HPLC?
* other method(s)?
I would like to know your comments/advices about my problem.
Thanks in advance.
INRA INA P-G Pathologie Vegetale 16 rue Claude Bernard
75231 PARIS cedex 05 FRANCE Tel +33 (1) 44.08.16.98 44.08.17.04
Fax: +33 (1) 44.08.17.00 Internet: bertheau at inapv.inapg.inra.fr
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