Fw: sequence services

Deanne Bell dbell at qnis.net
Wed May 9 16:36:20 EST 2001


O.K. Lets see if I am learning this stuff

Let's say I want to know the sequence from a DNA band obtained by doing a
RAPD.
I would need to clone it because 1)   I could not be sure that the band is
composed of the same fragment and 2)   both ends of the fragment would be
the same sequence (from the single primer). I would need to purify the band
from the agarose.  Then I would clone it into a vector with a known forward
primer sequence and a known reverse primer sequence.  Then select one
colony so that I know all the fragments in the sequencing are the same. 
Then purify that DNA -  with a plasmid mini-prep kit type of thing??  Then
send it to someone for sequencing.


BUT, let's say I have a DNA band from a PCR that specifically amplifies the
large sub-unit ribosomal DNA.  I can be pretty confidant that all the
fragments in that band are the same.  Then I could clean up DNA with a PCR
clean up kit and send it to someone for sequencing along with an aliquote
of my primer.

thanks for your time
Deanne Bell
Molecular Markers Lab Technician
USDA Agricultural Research Service
2021 S. Peach Ave.
Fresno, CA 93727
e-mail: dbell at qnis.net
phone: 559 453-3170
fax: 559 453-3088


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