Wisconsin-Madison/25 June 2009

June 25, 2009
Other: 

Reorganized -4 C fridge (plates) and -20 C freezer (samples) Advisor Meeting

Ex: Miniprep of Suspected BB NudF: 5 different colonies (A-E): 2 of each colony (10samples) Ran 0.7% Gel:

Cut with Xba Expected size:~2700bp

Results: Does not mean much …

Need better controls:

1.	BB cut nudF + backbone separate

2.	Backbone

3.	NudF

If inconclusive (several NudF’s have ligated together):

1.	one single cut for entire BioBrick inside NudF

2.	if multiple NudF’s = more than one bands

3.	THIS IS P2 (BB*) – DUMB FOOLS

Ex 15: Inducing various Modified (Triple) E.Coli Strain Cultures 3 Transformed Confirmed Triple DNA into following strains and plated:

1.	BL21 (DE3)

2.	K-12 MG1655 AaraBAD

3.	K-12 MG1655 (wildtype)

4.	DH10B

Ex 17: GFP Regulation by ProU Promoter

Test 2:

Redo Salt Tests GFP,ProU BB

'''Conc. / Culture (uL) / 5M NaCl (uL)'''

0.0 / 200 / -

0.2 / 192 / 8

0.4 / 184 / 16

0.6 / 176 / 24

0.8 / 168 / 32

1.0 / 160 / 40

Well Assignment:

'''Conc. /  Wells'''

0.0 / D5 / E8 / C6 / F9

0.2 / E5 / D8 / F6 / C5

0.4 / D6 / F4 / C8 / F8

0.6 / E9 / D5 / C7 / F7

0.8 / D9 / E4 / D7 / C4

1.0 / C9 / F5 / E6 / E8

All other wells were filled with MilliQ water

Ex 14: 

Digestion of Trip 1, Trip 2, MevT, MBI, NudF 18 uL RXN:

1.	12uL H2O

2.	 1.8uL Buffer 3

3.	0.18uL Protein BSA

4.	3uL DNA

Ran on 0.7% Agarose Gel

pNudF	4716 bp

pMevT	8593 bp

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

Results: We have triple transformation of interest

Ex 10: Cyanobacteria

Set up cyanobacteria apparatus

Inoculated PCC6803

Obtained Synechocystis sp PCC 6803 from Dr. Jenny Reed (inoculated 1 mL BG11 with 200uL of frozen stock) - NP