Experimental Procedure

Materials and Reagents The following materials and reagents were used throughout the experiment: P200 and P100 Pipetman micropipettes were used in this experiment. Fisher gel loading pipet tips were used. A Genesys 10S UV-VIS Spectrophotometer was used and had the ID number 2N3R294201. Two cuvettes (1.0cm path length) were used. A black Sharpie marker was used. A Fisher vortex mixer was used and had the ID number 060711069. A Fisher Scientific Isotemp was used and had the ID number 102NO562. Coomassie Brilliant Blue dye stain (2.5 g dye, 50% methanol, and 7.0% Acetic Acid in water) made on 4/22/2015 was used. Gel de-staining solution (50% Methanol/7.0% Acetic acid) made on 10/16/2013 was used. Two magnetic …show more content…
Six sheets of Parafilm "M" laboratory film was used and obtained from Neenah, WI. Gel Seal ¼ oz tube was used and had the ID number 80642143. A corks metric ruler was used. A black plastic wedge was used. A 60-cc and 10-cc syringe was used. Sample treatment buffer (STB) was used (prepared by adding 3.0 mL of glycerol, 0.1 g of ultrapure SDS, 6.0 mL of stacking gel buffer, 1.0 mL of -mercaptoethanol to a 15-mL conical centrifuge test tube and mixing by vortex action, and a pinch of Bromophenol blue dye). Three 10-mL and a 25-mL macropipette was used. Two 100-mL beakers were used. A 500-mL beaker was used. Stock slab acrylamide (30% = 29.2 g acrylamide/0.8 g bis-acrylamide per 100 mL of deionized water) made on 10/19/2015 was used. Upper tank buffer (prepared by dissolving 58.38 g of glycine, 12.0 g Triza base, and 4.0 g of …show more content…
The "cooling" device was connected to the bench water source via a hose and then the black electrode was plugged into the black socket and the red electrode was plugged into the red socket. The power supply was set on 20 mA until the tracking dye reached the running gel then the power went up to 35 mA to allow the gel to continue running until the dye was 2 cm from the end of the gel. Once the dye reached the end of the gel the power supply was turned off and the cords were disconnected. After removing the gel from the Hoefer SE 600 Ruby, it was stained with Coomassie Brilliant Blue for 4 hours then destained with SDS-PAGE Slab gel destaining solution for 16 hours to remove excess Coomassie Brilliant Blue. After destaining, the slab gel was then scanned by a Ricoh Super G3 gel