Team:UNIPV-Pavia/Calendar/August/settimana2

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(Difference between revisions)

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:

I21-1

I21-2

I21-3

I21-4

I21-5

I21-6

I21-7

I21-8

I21-9

I21-10

I21-11

I21-12

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

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

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.

@@ Line 37: / Line 37: @@
 |'''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
 |}
-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.
+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.
 ----

Team:UNIPV-Pavia/Calendar/August/settimana2

From 2010.igem.org

Revision as of 19:04, 9 August 2010

August, 9th

August, 10th

August, 11th

August, 12th

August, 13th

August, 14th

August, 15th