course-mutagenesis

Garry P. Larson glarson at ccmail.llu.edu
Mon Mar 13 19:07:51 EST 1995


Does anyone have any good ideas about a nice lab exercise for an advanced
Molec. Biol. lab for grad students in which you use site-directed
oligonucleotide mutagenesis (PCR or dut, ung ways) to mutate a gene.  The
goal of the lab is to expose students to oligo synthesis, mutagenesis,
sequencing, XF, and assay of phenotype of the mutants.  It would be great
if anyone had a "tried-and-true" protocol with vectors, strains etc.  The
ideal exp. would take 2-3 days from beginning to end (not counting the time
required to grow up the yeast XF's).  If somone had the appropriate
strains, vectors, and protocols, this would make things much easier. 
Perhaps a nice ts auxotrophic marker selection or a blue/white type of
screening.  Really, I'm open to suggestions.  If you can guide me to some
published lab protocols for a CSH class or equivalent I would be very
thankful

Please Email me with the scoop!!!!!
Thanks in advance,

Garry Larson
Ctr. for Molec. Biol. and Gene Therapy
Loma Linda Univ.
Loma Linda, CA 92350
V-(909)824-4300 x 81397
eamil glarson at ccmail.llu.edu



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