23 Jul WHAT CONTROL EXPERIMENTS WOULD YOU LIKE TO SEE HERE?
Gel Shift Assay. The interaction of a monomeric transcription factor (MW ~ 10000) with DNA is analy Show more Gel Shift Assay. The interaction of a monomeric transcription factor (MW ~ 10000) with DNA is analyzed by a get shift assay. In this assay a constant amount of 5-end 32p-labeled 20-mer duplex DNA oligonucleotide (double stranded DNA) is incubated with varying amounts of the transcription factor in 100 uL volume and the products run on a native polyacrylamide gel (the protein is not denatured so that you can detect and maintain protein-DNA complexes. An autoradiogram is made and the bands are exised from the gel and the radiation counts measured. A schematic drawing is shown below with the cpm found in each gel slice. a) what are bands 1 and 2? b) what is the Kd? c) What control experiments would you like to see here? Show less
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