Download Overview of Timeline for ES Cell Targeting and Southern Screening

Overview of Timeline for ES Cell Targeting and Southern Screening
Thaw ES cell frozen stock and culture
5 days

Electroporation of ES cells
24 hours

Start Drug Selection
8-10 days

Pick Colonies into 24-well plates
4-5 days

Freeze colonies into 96 well plates and start making genomic DNA from the 24 well
plates
5 days

ES cells are lysed; DNA is precipitated and resuspended in TE
24 hours

DNA is given to investigator to screen via Southern analysis for first positive
conformation
2 weeks

Investigator identifies positives and notifies TG facility of these
8 days

TG facility expands clones from 96 well plate to a T25, freezes the clones, and gives
investigator DNA from these clones for second positive confirmation
2 weeks

Investigator re-confirms positive clones and notifies TG facility

TG facility can begin blastocyst microinjections of modified ES cells, or can perform
additional electroporations on these cell lines if needed
Total estimated time: 35 days
 This does not include time taken by an investigator’s lab to perform Southern
analysis for conformation of positive clones
 Also, due to the growth rate of cells, and differences in drug selection times, this
timeline may be shortened or lengthened a bit