1. iGEM Bioware
Week 9

2. Arsenic Bioremediation
Got ArsR into plasmid backbone and began site directed mutagenesis. 
using NEB competent cells significantly improved efficiency.
PCR of ArsB and LamB is still not working--Any suggestions?  Both these proteins are vital to our project. 
Ordered new primers for RpoS promoter and ArsR Promoter with EcoRI cut site to increase yield and efficiency.
Also worked to get constitutive promoters in front of gas vesicles biobrick but getting lawns after transformations. 

3. Expected Sizes: pRpoS438: 438 bp pRpoS232: 232 bp ArsB: 1290 bp LamBF1R: ~1.3 kb LamBF2R: ~1.3 kb ArsR fusion: ~350 bp ArsR: 354 bp pArsRF1R1: 540 bp pArsRF1R2: 547 bp pArsRF2R1: 293 bp pArsRF2R2: 300 bp
ArsB
LamB
F2R
pRpoS
438
ArsR
pArsR
F1R1
pArsR
F1R2
pRpoS
232
LamB
F1R
ArsR 
Fusion
~500 bp
pArsR
F2R1
pArsR
F2R2
(-)

4. (-) Expected Size: ArsR insert: ~600 bp ArsR 1 ArsR 2 1 kb ladder
ArsR 1
ArsR 2
(-)
1 kb
ladder
Expected Size:
ArsR insert: ~600 bp
500 bp

5. ArsR in Backbone
Transformation efficiency highly improved by using NEB cells instead of our own competent cells

6. Gold Finally got golT out of Salmonella after using genomic prep kit
Still haven't been able to get gesABC
going to order primers for gesABC proteins individually instead of trying to get the entire operon
Did a digestion/ligation yesterday of golT, golB and golS
going to transform and plate them Monday since we can't come in during the weekend

7. Gel of PCR Expected Sizes: golT = 2300bp golB = 200bp golS = 500bp
gesABC = 6000bp

8. PCR - genomic and PCR purification template
We keep getting weird bands for gesABC and aren't getting the correct 6kb band, so we are going to order primers for ABC individually

9. Digestions - XbaI + SpeI

10. Decoder 
Update
Transformation efficiencies increased significantly using NEB 5a and Turbo electrocompetent cells.
Still searching for Gen5 software for HT Synergy Plate Reader
Matt is working on 
putting together a light 
cannon

15. Dippin' Dots Fundraiser
We had good sales Wednesday despite the rain, selling 45 cups!
Selling again this afternoon from 12:30-3:30 in front of the Union