IGEM:University of East Anglia (UEA), Norwich, UK/2009/Notebook/NRP-UEA-Norwich iGEM/2013/07/10
|Week Six||Main project page|
Previous entry Next entry
Transformation plates were retrieved, Three colonies from each of the two plates (2a,b,c and 3a,b,c) were inoculated into a culture and put onto incubation overnight at 37°C with vigorous shaking.
DNA was isolated, by performing minipreps, from the two cultures that contain standard E.coli cells. This yielded two samples, one from each culture, which were stored in the -20°C freezer. The remaining two cultures that contain BL21 E.coli cells were used for protein expression. This was to be performed in stages over the next few days. Firstly two aliquots of 100μL were taken from each culture (1,2) and added to seperate 10ml LB solutions containing Kanamycin (10μL) and Chlorophenicol (15μL) respectively. These four new cultures (1a,1b,2a,2b) were left to grow at 37°C in a shaking incubator for 2 hours until an absorbance between 0.4-0.6 at OD600 was shown after testing with a Biophotometer. 40μL of IPTG was added to half the cultures (2b,1b) All four cultures were left to incubate overnight at 25°C.
In the team meeting we discussed how to record data about soil samples we receive. It was decided the best approach would be to set up a spread sheet and issue the tokens with a soil number. This meant, at a later date, the initial testing results from the samples could be put on our blog so and people could check their sample using the issued number. With the meet up only a few days away, we practiced the presentation a few times. The poster and soil token cards also arrived today