Team:UNIPV-Pavia/Calendar/August/settimana2

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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
|'''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.
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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-run and gel-extracted right amplicons.
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Revision as of 19:04, 9 August 2010


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:I21-1I21-2I21-3I21-4I21-5I21-6I21-7I21-8I21-9I21-10I21-11I21-12
I21-new:I20-1I20-2I20-3I20-4I20-5I20-6I20-7I20-8I20-9I20-10I20-11I20-12I20-13I20-14I20-15I20-16I20-17

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-run 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

Than we streaked cultures on a six-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 (without the right resistance) 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