Wednesday, December 3rd, 2008
Hopefully you're one of the first 100 researchers who have ordered vectors, ES cells, and/or mice from us...as we've just passed the 100 orders milestone!
So on this occasion, I'd like to take this opportunity to get your feedback on how we're serving you...timelineness, responsiveness, technical service...anything you'd like to inform us about.
Edward Kelly said...
I must say I am glad to have KOMP clones available and I am making mice for two different gene knockouts. The one minor complaint I have is the genotyping protocols. For each targeted allele, PCR primers have been designed as part of the QC but I have not been able to get their sequences for either CSD or Regeneron clones. These should be included as part of the service.
January 8, 2009 9:07 AM
Brandon Willis said...
Dear Kelly, the sequences are available by entering the gene name in the search field of KOMP.org and then click on gene name which then shows all projects for the gene targeting. click on the project ID relating to your order (e.g. CSD30462 for Cyp4b1 or VG10578 for Cyp4v3) and this will show the targeting vector map along with primer and vector sequences. i hope this is helpful.
January 9, 2009 8:35 AM
julia Dorin said...
Is there an easy protocol for carrying out the DT negative selection after electroporation into ES cells?
August 27, 2009 4:12 AM
Bryant Gavino said...
One of the advantages of DT over TK is that it doesn't require any drug selection. If DT is integrated in a random insertion event, this will be lethal to the cell. As described in Yagi et al. (Analytical Biochemistry 1993), DT is composed of fragments A and B. The B fragment participates in binding to the receptor on the cell surface and facilitating the transfer of the A fragment to the cytoplasm. The A fragment catalyzes the transfer of the ADP-ribosyl group of NAD+ to elongation factor (EF-2) in the cytoplasm, thereby inactivating EF-2, inhibiting protein synthesis, and killing the cells.
KOMP’s work has shown that DT is much more efficient over TK for negative selection.
August 27, 2009 12:10 PM