Team:Paris/30 July 2009

NoteBook
July 30th

 

Lab work
 Reception of strains
 * New DH5&alpha;
 * DH5&alpha; with g3p in plasmid

 Glycerol stock of new DH5&alpha; [S6]
 * Stock in 20% glycerol at 80°C

 New competent DH5&alpha; by RbCl
 * Competent bacteria
 * 1/50 ON culture of New DH5&alpha;
 * OD600=0,6
 * Transformation with puc19 (10pg/µl)
 * 250µl of RbCl competent DH5&alpha;
 * 12,5µl puc19
 * Leave 20 min in ice
 * 45s at 42°C
 * Leave 2 min in ice
 * Add 1ml of hot LB (42°C)
 * 1h at 37 under agitation
 * Put on plate (LB agar + Amp)

 Overnight culture for [S3] &Delta;nlp1, [S4] TB28, [S5] MG4, [S6] DH5&alpha;, [S7] DH5&alpha; g3p
 * S3 &Delta;nlp1: 5ml LB + 5µl Kan 1000x
 * S4 TB28 : 5ml LB
 * S5 MG4: 5ml LB + 5µl Kan 1000x + 250µl NaCl 20%
 * S6 DH5&alpha;: 5ml LB
 * S7 DH5&alpha; g3p: 6ml LB + 6µl Amp 1000x

 Stock of NaCl 20%
 * 100ml of H2O
 * 20g of NaCl
 * Filtration

 