Wisconsin-Madison/22 June 2009

June 22, 2009
Other: Primer design of pmmBraA Plan to have psbA1 synthesized for ~$50 with biobrick sites (psbA2 conj has biobrick sites within its sequence

Ex 10: Cyanos are alive – good sign Plated Transformed Cyanobacteria of Cm plates Made freezer stocks of PCC 7942 to test the e coli protocol as a viable method for cyanobacteria preservation Inoculated new PCC 7942 stocks from 6/10 stock which are a deep green - NP Tried simple transformation of PCC 7942 again with four different concentrations/plates (50uL culture+1uL pMMBRraA)(50uL each of 2:1, 	3:1, 1:1 concentration increases on CM+BG11 plates and 50uL of 1:1 on BG11 plate) - NP

Ex 16 F: growth of DH10 B with:

1.	Control - vector only - Undigested = lawn (red) = OUR DH10B cells are very competent

2.	Choline = no growth

3.	Pro U = 100 colonies – ONE WHITE COLONY! (Biobrick)

4.	ProU + GFP = 2 colonies (Biobrick)

5.	YhfR = no growth

6.	Nud F = no growth

Both iGEM Plasmid: pSB1A3 different digestions

P1 - cut Ecor1and Spe1

P2 - cut Xba1 and Pst1

G. Streak out separate plates of pink colored colonies on ProU (unequal division) -grew all pink

Ex 14: Digestion of Trip 1 and Trip 2 18 uL RXN:

1.	12uL H2O

2.	1.8uL Buffer 3

3.	0.18uL Protein BSA

4.	3uL DNA

5.	1ul Enzyme BamH1

Ran on 0.7% Agarose Gel: pic



1Kb Ladder

pNudF 4716 bp

pMevT 8593 bp

pMBI 6000 bp --> (cut twice: 4000, 2000)

Ex : Transform P1 and P2 which are backbones to get control for just ligation according to transformation protocol

Ex : Inoculate liquid cultures of

1.	gsSDMT

2.	backbone

Ex 10: Cyanobacteria

Made freezer stocks of PCC 7942 to test the e coli protocol as a viable method for cyanobacteria preservation - NP - inoculated new PCC 7942 stocks from 6/10/09 stock which are a deep green - NP - tried simple transformation of PCC 7942 again with four different concentrations/plates (50uL culture+1uL pMMBRraA)(50uL each of 2:1, 3:1, 1:1 concentration increases on CM+BG11 plates and 50uL of 1:1 on BG11 plate) - NP