Stu Cain (s.a.cain at sheffield.ac.uk) wrote:
: In article <CwIDvH.BuE at murdoch.acc.Virginia.EDU>, vlm4b at darwin.clas.Virginia.EDU (Vicky L. Miller) says:
: >Dear fellow Pichia protein expressers,
: > This is not good news. It has taken me months to get my
: >Dna into one of the secretion vectors. Tonight I plated from
: >RDB plates onto the MM and MD plates. I'll let you know what
: >happens. What concentration of antibiotic do you use in your
: >Vicky Miller
: >Graduate Student whose life hangs in the balance
: Dear Vicky
: I have found there is no need to add any antibiotics
: to any of your plates or media as long as they are prepared
: reasonably sterile.
: Good Luck with your selection.
: Stu Cain
: Another graduate student whose at the mercy of Pichia pastoris
Dear Stu and Vicky,
I haven't had a lot of problems getting my DNA into the pHIL-S1 expression
vector. The problem is that the yeast will not express what I put in
there. I have been at this for a good 6 months. I have now decided to
use one of the other vectors--I am having problems there. I think
Invitrogen isn't telling us everything there is to know because I know of
too many people who have been at this for a year and still have no results.
A third graduate student whose at the mercy of Pichia Pastoris