We began our project by preparing material needed. We made liquid and solid medium using LB powder. We also made solution for transformation process like MgCl2, and TSS medium. In this week, we also culture the stock of our RFP in liquid medium. We aim to make the stock of RFP and also examine whether the RFP still work or not. The culture of RFP is not growth. It indicates that our RFP stock has been damaged due to the unstable storage temperature.

Detail Activity:

1.August 14 th, 2014

Making liquid medium using LB medium powder, sterilization using autoclave, add antibiotic, culture E coli DH5α, shaking

2.August 15th, 2014

Making solution MgSO4, glucose 1 M, CaCl2, TSS medium

3.August 16th, 2014

Making solid, liquid, and NA medium, Culture E coli (Glycerol stock RFP)

4.August 17th, 2014

Add chlorampenichol to solid medium (LB medium), streak RFP (stock 6 tubes), check culture E coli (Glycerol stock RFP)

Week 2

In the second week we culture the stock of RFP into the solid medium. The purpose is to make the stock of RFP in the solid medium. We also transformed the plasmid Bba_J04450 and Bba_B0034 into E coli BL21 (DE3). We did transformation twice in this week. The first transformation does not show any red colony growing. But the second transformation showed red colonies growing indicates that the transformation process success.

Detail Activity :

1. August 18th, 2014

Making solid medium with chlorampenicol, take Bba_B0034 from kit plate and storage

2. August 19th, 2014

Making ampicilin stock 1 M, streak RFP (stock lab) medium padat, transformation

3. August 20th, 2014

Check Bba_J04450 transformation result, culture RFP, streak RFP, making ampicilin stock 1 M, plating transformation result, streak RFP from solid medium

4. August 21th, 2014

Check transformation results (Not growing), check RFP streak (contaminant), streak transformation result

5. August 22th, 2014

Culture E coli glycerol stock (BL21(DE3)), Transformation

Week 3

Detail Activity

1. 25 August, 2014

Culture RFP DH5α (Add over expression reagent), making chlorampenicol stock

2. 26 August, 2014

Culture E coli glycerol stock RFP, culture over expression DH5α in 6 tube, streak RFP from single colony and line colony to the LB medium with and without chlorampenicol

3. 27 August, 2014

Plating transformation results, streak stock RFP