Team:Wageningen UR/Notebook/Proj1/June

June 1

12 colonies were screened by colony PCR. A BBa_10462 transformant was used as a positive control and plasmids of BBa_10462 were used in the PCR.

The PCR was confirmed for some of the samples. The corresponding colonies (BBa_10462, -K325210, -E0422, -C0062 and -P0412 transformants) inoculated 10 mL LB medium with Amp and were incubated on a shaker at 37°C overnight.

June 2

With colony PCRs, transformed colonies were found with: BBa_1712019, -"K325210" and -K325219. No transformed colonies were found that should contain: BBa_R004, -K325909, -R0062 or -R0063. The former colonies were grown in 10 mL LB medium in presence of Amp on a shaker at 37°C overnight.

All inoculations of June 1, but one: the BBa_K325210 transformants, yielded enough growth. The specific colony was incubated, in the same way, again. 5 mL of each of the other cultures was used in miniprepping.

June 3

Of all incubations, only cultures of BBa_1712019 transformants were made. These were plated on LB medium with Amp.

The yield of miniprepping was determined by using the Nanodrop spectrophotometer. The plasmids were present in the following concentrations:

• 95.4 ng/µL of BBa_C0062;
• 61.0 ng/µL and 70.5 ng/µL of BBa_E0422;
• 86.2 ng/µL of BBa_I0462 and
• 86.2 ng/µL of BBa_P0412.

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