Team:Newcastle/Labwork/12 October 2009

=Formal Lab Session - 12th October 2009=

Pouring the plates
Today we poured plates for our B. subtilis transformations

For CHL: Use: 5ug/ml For Em:use 0.3ug/ml

Our stocks are: Em Stock: 1mg/ml CHL stock:2.5 mg/1ml
 * 500ml LB + 150ul of Em
 * 1500ml LB + 450ul Em + 3ml CHL
 * 500ml LB + 150ul Em + 1000ul CHL + 5gr Starch (Use 10gr Starch for a litre and add the starch before sending it for autoclaving, antibiotics are after the autoclave!)
 * 500ml LB + 150ul Em + 1000ul CHL + 5gr Starch + 500ul IPTG(1mM)

pMutin4 plating out
Pmutin4 plate we prepared from last Friday did not work. We plated them out from frozen stock.

Sending parts to the registry
Measured the concentration of 1:10 diluted primers for pSB1AT3 Forward primer: 185ng/ul (30pmol/ul) Reverse primer: 245 ng/ul (40pmol/ul)

Culture 3 B.subtilis strains which had been integrated in pMutin4 vector
BFS 40 BFS 867 BFS 2426


 * Plate these 3 strains on LB+Em plate

Culture 3 strains for B.subtilis transformation

 * Followed the B.subtilis transformation protocol from our lab protocal.