On Tuesday this week, we prepared 12 tubes with 5 mL LB liquid media/ 5mcL kanamycin.
On Wednesday, we innoculated 6 random colonies from Plate B (350bp fragment), and 6 random colonies from Plate F(450bp fragment), each in seperate tubes prepared Tuesday. We allowed these to shake at 200 RPM/ 37°C overnight.
On Thursday, we isolated plasimd DNA from each of the tubes innoculated on Wednesday. Glycerol stocks were then created from the remaining cultures for each. We then digested each mini-prep with PstI and XbaI enzymes to release our inserted fragment. These were allowed to run on a 0.8% agarose gel to determine whether the transformation from Nov. 15th was successful.