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From 2012.igem.org

Week 1

6/07/2012

Wet Lab

Today we transformed both versions of cph8 from 2012 Distribution.

Dry Lab

We also placed order for the following new BioBricks:

• BBa_K207001 - PhyB-DBD Fusion
• BBa_K422012 - Pif3 (Aar1 C part)
• BBa_K422013 - PhyB (Aar1 C part)
• BBa_K592006 - pFixK2
• BBa_K592004 - YF1
• BBa_K592005 - FixJ
• BBa_K592000 - Cph8
• BBa_K365000 - Pif3

In addition, we brainstormed and developed a new idea to use the PYP/GCN system to activate a gene.

6/08/2012

Wet Lab

No colonies appeared in the transformation of both cph8 a/b. We suspect that the failure of the transformatiosns were due to bad sequences.

Week 2

6/11/2012

Wet Lab

We performed PCR to obtain mCherry from the NAS 151 plasmid. Following this we ran a 1% gel and purified the product which resulted in a 30ns/λ yield.

6/12/2012

Wet Lab

We digested mCherry PCR product with restriction enzymes BamHI and NotI. Then we column purified mCherry and ligated into NAS152 backbone. Afterwards we transformed NAS152-mCherry into DH5alpha cell lines. This was then poured 25 LB-Kan plates.

6/13/2012

Dry Lab

Read papers to research the project.

6/14/2012

Dry Lab

Continued to read papers.

6/15/2012

Dry Lab

Met with Mark Goulian to develop the project. Also obtained pDawn and pDusk plasmids.

Week 3

6/18/2012

Wet Lab Did a test cut of pDawn and pDusk plasmids using Xma1. Ran an analytical gel. Then cut the two plasmids for ligation using BamHI and NotI . Finally ran and purified the gel.

Dry Lab We ordered and picked up PCR purification kit. Additional components were also ordered.