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Methods for quickly determining the chromosome number present in targeted mESC lines.

Keywords: mESC karyotyping

Category
Protocol
Record History
Added on April 13, 2010 at 2:59 PM by Skelton, Jennifer
Modified on August 14, 2013 at 1:32 PM by Administrator, Labnodes
Shared with (contributions)
TMESCSR: Protocols

Reagents. supplies, and equipment:

  • cES cell media
  • 60 mm tissue culture dish with MEFs

Procedure:

  1. Submit the service request form for karyotyping.  http://labnodes.vanderbilt.edu/resource/view/id/3057
  2. The ESC Shared Resource will schedule and coordinate a time for cells to be delivered for karyotyping.
  3. E-mail the names and clone numbers of the cell lines requested for karyotyping to Jennifer Skelton (jennifer.skelton@vanderbilt.edu).  Label the culture dishes with the same names.
  4. On a 60 mm culture dish, plate cells on a layer of MEFs.
  5. Colonies need to be at least medium in size and 50% to 90% confluent for karyotyping.  This is usually about two days of growth.
  6. On the day dishes are being handed off to the TMESCSR for karyotyping, feed the plate(s) with cES before 10:00 AM and communicate to the TMESCSR personnel what time they were fed.
  7. Bring plates for karyotyping to the TMESCSR in 9410 MRB IV by 11:30 AM.

Source: TMESCSR 2/13