SOS Call to all experinced

martin LEACH leach at bu.edu
Tue Apr 2 08:05:19 EST 1996


IMHO...

get some T-vector....make it or buy it
clone 'em and sequence 'em

Sometimes doing the old fashioned cloning works better than getting a poor
cycle
sequencing result

Martin

Daniel Pereira (mgs6145 at student.health.ufl.edu) wrote:
: spyong at pc.jaring.my wrote:
: >
: > Hello,,
: > 
: > The only sequencing I've ever done is through the text book. However, it
: > seems like now is the time to start sequencing. I have three PCR products
: > and need to sequence them before I work further only to find that they are
: > junk. Someone suggested end-labbeling, others say double strand sequencing,
: > some say cycle sequencing....some say stick to 'Sequanase' (whatever that
: > means). Do I have to subclone into M13?
: > 
: > Please,I know sequencing is not easy and may be take ages, but I just need
: > to know which would be to best method and time saving. My bands are
: > 200bp,350bp and 600bps long.
: > 
: > Thanks a million.
: > 
: > My e-mail : spyong at pc.jaring.my
: > 
: > Cheers,
: > Stephanie
: > 
: >  
: > 
: If the terminal 40 bp or so are expendable I would suggest using the
: Boeringer Mann. kit for cycle sequencing.  You can use either of your 
: PCR primers and it is simple, and efficient.  Best of Luck
: Dan

--
.....          Martin Leach                Email:leach at bu.edu
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329
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