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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.
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
Than we streaked cultures on a four-sector divided plate. Since 6 ug/ml is a very low concentration, we wanted to check if actually nothing (without the right resistance) grow on these plates.
Transformation of 100 ul of MG008 and MC123 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.
August, 10th
August, 11th
August, 12th
August, 13th
August, 14th
August, 15th
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