Team:UNIPV-Pavia/Notebook/Week16

From 2008.igem.org

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*Colony PCR for the two ligation plates:
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*Colony PCR for the two ligation plates
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{|
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|[[Image:pv_colonypcr_30_35again.jpg|thumb|370px|left|Marker 1Kb, Lig.30 (5 colonies), Marker 1Kb, Lig.35 (5 colonies)]]
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|}
*Gel results:
*Gel results:

Revision as of 17:49, 2 October 2008


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Notebook



Week 1 Week 2 Week 3 Week 4 Week 5 Week 6 Week 7
Week 8 Week 9 Week 10 Week 11 Week 12 Week 13 Week 14
Week 15 Week 16 Week 17 Week 18 Week 19 Week 20 Week 21



Week 16: 09/1/08 - 09/5/08

09/1/08

  • We had to perform these ligations:
    • Lig.22-R0062
    • Lig.31-Lig.22
  • but we didn't have enough DNA for 22 and 31 plasmids...
  • We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.

09/2/08

  • Glycerol stocks/miniprep for Lig.22 and Lig.31.
  • Plasmid digestion for:
    • Lig.22 (E-S)
    • Lig.22 (X-P)
    • Lig.31 (S-P)
    • R0062 (E-X)
  • Run/gel extraction.
  • Ligations:
    • Lig.31-Lig.22 (=Lig.35)
    • Lig.22-R0062 (=Lig.30)
  • We incubated ligations at 16°C overnight.

09/3/08

  • We transformed/plated ligations.

09/4/08

  • Colony PCR for the two ligation plates
Marker 1Kb, Lig.30 (5 colonies), Marker 1Kb, Lig.35 (5 colonies)
  • Gel results:
    • Lig.30 (3rd colony)
    • Lig.35 (gel showed an unexpected contaminant...we decided to re-perform this ligation)

09/5/08

  • Glycerol stock/miniprep for Lig.30.
  • We sent Lig.30 (3rd col) to Primm for sequencing.