Week 5:
University of Iowa Department of Biochemistry

June 18, 2013

The purified plasmid containing the FLAG-tagged FANCJ was run on an agarose gel to confirm the proper vector and insert were present. The Gel showed that they were.

Following the confirmation of FANCJ in the purified plasmid, the cultures of mammalian cells, HEK293T, were transfected and left to incubate overnight.  The protein from the transfected cells was purified the same as before but with an extra gel chromatography purification step after the crude filtration.  Some of the eluate from the crude filtration was saved to be analyzed for FANCJ content along with the fractions from the gel chromatography.