IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/07/30

{| width="800"
 * style="background-color: #800000"|[[Image:UIUC_ArsR_System.jpg|150px]] UIUC - Arsenic Collection System!
 * style="background-color: #AFEEEE" align="center"|  |Main project page
 * style="background-color: #AFEEEE" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

7/30/10
8:20 AM

checked 5 mL culture overnights, 2 were green, 2 were normal colored.

Made glycerol stock of LamBF2R: .9 mL 50% glycerol, .9 mL culture.

8:30 AM

PCR Cleanup

promega kit

pArsRF1R2: 16 ng/μL

pRpoS438: 16 ng/μL

8:35 PM

Miniprep

Qiagen kit

LamBF2R: 68 ng/μL, 20 μL left

11:55 AM

Digestion

pRpoS438:

pArsRF1R2

pSB1C3

incubate at 37°C for 15 min, inactivate at 80°C for 20 min

12:40 PM

Ligations

pRpoS438-pSB1C3

pArsRF1R2-pSB1C3 incubate at room temperature for 10 min, inactivate at 80°C for 20 min.

12:45 PM

set up 5 mL cultures of ArsR-B0015-pSB1A2

1:30 PM

Transformations

100 μL cells, 3 μL ligation product. electroporate, add 1 mL SOC, recover for 1 hour at 37°C

Plated 200 μL cells and 75 μL cells on CAP plates for each sample at 2:30 PM.

1:45 PM

Miniprep

Qiagen kit

Green LamBF2R: 285 ng/μL

Forgot to make glycerol stock.


 * }