Matt Gethers/CRI, Thailand/Labwork/Binding Assay/8.7.08 Run Notes

=8.7.08 Binding Assay Run=

Ran experiment testing GpxR binding affinity for both the HmgA and Gpx1 promoter regions. Got 10x binding buffer from p'So and made dilutions of lot 1 of GpxR as follows:

Made ~18 &mu;l of 10x Buffer+DTT using 16.3 &mu;l 10x and 1.63 &mu;l 100 mM DTT. Diluted as follows:

Placed dilutions on ice, then to an FA cuvette added: 900 μl MilliQ water, 100 μl 10x buffer, 1 μl 0.5 mg/ml polydIdC, and 1 μl 1 M DTT. To first cuvette, added 1 &mu;l 10 &mu;M Gpx1 promoter probe (final concentration of 10 nm). Added the same amount of HmgA promoter to second cuvette which otherwise had all the same reagents in the volumes stated above.

Data
August 8, 2008 GpxR Data