Lab Notebook

6 August 2009

Sriram

Today I continued with the ligation of the restricted parts done yesterday. Then the ligated parts were electro transformed and also heat shock transformed. There are in total 8 assembly as follows: 1. pTet + RBS-cI-RBS (Chloramphenicol backbone)
2. mRFP + Double Terminator (Chloramphenicol backbone)
3. pLacI + RBS (Chloramphenicol backbone)
4. TetR + Double Terminator (Chloramphenicol backbone)
5. pTet + Lock3c (Chloramphenicol backbone)
6. cI + Double Terminator (Chloramphenicol backbone)
7. λp-in + RBS-GFP-Double Term (Chloramphenicol backbone)
8. pLacI + key3c (Amp-Kan backbone)

Calin

Good results on the conjugation test. Many colonies. All plates were imaged on the safe imager with the 5 Megapixel camera. The plates with GFP (T and R) were excited by blue light and placed under a filter (50 ms exposure). The plates without GFP (D) were imaged with the upper white light and an exposure of 15ms. Images were color inverted before the image analysis. Colony counting was done with [http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=1770926 Clono-Counter].

Calculated conjugation efficiencies:

Trimethoprim control plate had no growth.

5mL tube of assembly CA made.

Elecroporated 10 assemblies. Experienced some arcing with assemblies 1 and 6. Assembly 6 was redone with only 2 uL of DNA added.

Tim Weenink

Elecroporated 10 assemblies.

Orr

Made 2 one liter batches of LB medium, and prepared some chemically competent TSS cells.