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AUGUST: WEEK 2
August, 9th
Phasins plates showed very few colonies (12 for I20-new and 17 for I21-new): they were all picked and let grow in LB+Amp 100ug/ml. In the evening we made glycerol stocks and re-filled the tubes for the screening of the following day:
| I20-new: | I21-1 | I21-2 | I21-3 | I21-4 | I21-5 | I21-6 | I21-7 | I21-8 | I21-9 | I21-10 | I21-11 | I21-12
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| I21-new: | I20-1 | I20-2 | I20-3 | I20-4 | I20-5 | I20-6 | I20-7 | I20-8 | I20-9 | I20-10 | I20-11 | I20-12 | I20-13 | I20-14 | I20-15 | I20-16 | I20-17
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In order not to loose our time we decided to perform again the PCR-amplification/modification of <partinfo>BBa_K208001</partinfo> in case of a negative screening. Than we gel-ran and gel-extracted right amplicons.
Check of LB+Cm 6 ug/ml agar plates. We let grow at 30°C:
- MG1655 in LB
- MC1061 in LB
- MG123/008 in LB+Amp 50 ug/ml
- MC123/008 in LB+Amp 50 ug/ml
- F2620-4C5 (positive control)
Than we streaked cultures on a seven-sector divided plate, and we let grow it ON at 30°C. Since 6 ug/ml is a very low concentration, we wanted to check if actually nothing (except F2620-4C5) grow on these plates.
Transformation at 30°C of 100 ul of MG13/008 and MC123/008 competent cells to check their efficiency. We used 1 ul (~4 ng) of RING, F2620-4C5, Nothing (water) and plated on proper LB agar plates:
- RING: Cm 34 ug/ml
- F2620-4C5 (positive control): Cm 12,5 ug/ml
- Nothing (negative control): Cm 12,5 ug/ml
Both RING and F2620-4C5 should survive, but we had a problem so that MC008 transformed with RING couldn't be plated. We will check it another time. We let grow plates ON, 30°C.
August, 10th
August, 11th
August, 12th
August, 13th
August, 14th
August, 15th
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