User:Mbennie/Notebook/Lab Notebook/Notebook/2007/08/01

Cellular Adhesion

 * Remaining oligos arrived today:
 * IgAb-F
 * IgAb-R
 * IgAss-F
 * IgAss-R
 * Fos-F
 * Fos-R
 * JunB-F
 * JUnB-R
 * GCN4-F
 * GCN4-R
 * Resuspended in TE to 50uM


 * PCR
 * Performed PCR on N. gonorrhea and BBa_C2001
 * Tubes with 40ul PCR Supermix, .4ul N. gonorrhea DNA, and .4ul of each primer:
 * A: Signal Sequence IgAss-F and IgAss-R
 * 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
 * Tubes with 40ul PCR Supermix, 1ul cell dilution (picked colony diluted in 100ul water), and .4ul of each primer:
 * F: GCN4 Leucine Zipper GCN4-F and GCN4-R
 * PCR protocol
 * 5mins@95C
 * 20secs@94C
 * 20secs@55C
 * 1min@68C
 * REPEAT 2-4, 34 times
 * 5mins@70C
 * FOREVER@4C


 * Gel
 * Ran 1% gel for 40 minutes at 100V with samples (5ul sample with 2ul of loading dye)
 * BANDS APPEAR TO BE THE RIGHT SIZE!!!
 * The colony PCR did not work as well as the others