User:Mbennie/Notebook/Lab Notebook/Notebook/2007/08/11

Cellular Adhesion

 * Glycerol
 * Took 1.6ml of liquid culture and created 10% glycerol stocks
 * IgAbc #8


 * Miniprep
 * Extracted DNA from remaining liquid culture (~6ml)
 * Eluted in 50ul water


 * Liquid culture
 * Grew up cultures in 8ml Amp/Kan LB (didn't have mixed, so used 4ml of LB KAN and 4ml of LB Amp) inoculated from plates
 * IgAss + Fos #1
 * IgAss + Fos #2
 * IgAss + Fos #3
 * IgAss + JunB #1
 * IgAss + JunB #2
 * IgAss + JunB #3
 * Grew up cultures in 8ml Amp/Cl LB inoculated from plates
 * GCN4 #4
 * GCN4 #5
 * GCN4 #6
 * PCR
 * Template: 40ul PCR Supermix, .4ul N. gonorrhea DNA, and .4ul of each primer
 * B: Part 1 of IgA beta-core IgAb-F and Mut_Pst1a-R
 * C: Part 2 of IgA beta-core Mut_Pst1a-F and Mut_Pst1b-R
 * D: Part 3 of IgA beta-core Mut_Pst1b-F and IgAb-R
 * E: Full IgA beta-core IgAb-F and IgAb-R
 * Same thermocycler protocol as 8.1.2007


 * PCR
 * Template: 40ul PCR Supermix, .4ul VF2 and VR, 1ul IgAbc #8 miniprep
 * Same thermocycler protocol as 8.1.2007


 * Gel
 * IgAbc #8 looks correct
 * Try sequencing it again