Team:Newcastle/Labwork/13 October 2009

=Formal Lab Session - 13th October 2009=


 * Today we decided not to characterize cotC+smtA biobricks since we won't have time to finish and we want to finish the lab work this week.
 * We will do B.subtilis transformation using 3 strains (BFS867, BFS40, BFS2426) cultured yesterday.



Preparation of parts to send to the registry
Measured the concentrations of the midipreps and prepared them to send to the parst registry sspB:64 ng/ul To parts registry: 6.4 ng/ul in 20ul

ara:34 ng/ul To parts registry: 3.4 ng/ul in 20ul

ara+sspB:87.5 ng/ul To parts registry: 8.75 ng/ul in 20ul

cotC+smtA:355.9 ng/ul To parts registry: 9 ng/ul in 20ul

cwlJ:76.9 ng/ul To parts registry: 7.69 ng/ul in 20ul

sleB:98.3 ng/ul To parts registry: 5 ng/ul in 20ul

swlJ + sleB:55 ng/ul To parts registry: 5.5 ng/ul in 20ul

Preparation of parts for sequencing
Prepared the primers for psB1AT3 (x2, prepared two sets)

Forward primer preparation: Total 30ul, 10pmol/ul Forward primer : 10ul (30pmol/ul) PCR water: 20ul

Reverse primer preparation: Total 30ul, 10pmol/ul Forward primer : 7.5ul (40pmol/ul) PCR water: 22.5ul

Placed into the box for sequencing parts

B.subtilis transformation

 * Followed the standard protocol form our lab's B.subltilis 168 transformation protocal.
 * Added 5ul pGFP-rrnB:kinA vector to all three strains.
 * Plated the transformed cell on LB+Em+CHL plate