Team:UNIPV-Pavia/Notebook/Week15

From 2008.igem.org

(Difference between revisions)
Line 84: Line 84:
*Colony PCR for 5 colonies of Lig.12-'''B1006''' (called Lig.22).
*Colony PCR for 5 colonies of Lig.12-'''B1006''' (called Lig.22).
 +
 +
{|
 +
|[[Image:pv_colonypcr_22again.jpg|thumb|370px|left|Lig.22 (5 colonies), Marker 1Kb]]
 +
|}
*Gel results: we chose 1st colony to grow a 9 ml overnight culture.
*Gel results: we chose 1st colony to grow a 9 ml overnight culture.

Revision as of 13:27, 2 October 2008


Home.jpg Home Unipv logo.jpg The Team And.jpg The Project Safety.jpg Biological Safety Dna.png Parts Submitted to the Registry
Laptop.jpg Dry Lab Pipette.jpg Wet Lab Math.gif Modeling Note.jpg Protocols Notebook.gif Activity Notebook



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 15: 08/25/08 - 08/29/08

08/25/08

  • Pladmid digestion for:
    • R0051 (S-P)
    • R0040 (S-P)
    • B0030-C0061-B1006-R0062-B0030-E0040-B1006 (E-X) (=Lig.b (E-X))
    • B1006 (E-X)
    • Lig.12 (E-S)
  • Run/gel extraction.
  • Ligations:
    • BBa_R0051 (S-P) - BBa_E0240 (X-P) (for promoter test)
    • BBa_R0040 (S-P) - BBa_E0240 (X-P) (for promoter test)
    • Lig.12 (E-S) - B0030-C0061-B1006-R0062-B0030-E0040-B1006 (E-X) (for AND logic gate test)
    • Lig.12 (E-S) - BBa_B1006 (E-X) (to re-perform mutated assemblies)
  • We incubated ligations at 16°C overnight.
  • We ordered 3OC6HSL (Sigma).

08/26/08

  • We transformed/plated ligations.

08/27/08

  • Plates grew correctly. We checked colony fluorescence of three plates under UV rays:
    • R0051-E0240 glowed
    • R0040-E0240 glowed
    • Lig.12-Lig.b (R0051-B0030-C0062-B0030-C0061-B1006-R0062-B0030-E0040-B1006) glowed
  • Colony PCR for 5 colonies of Lig.12-B1006 (called Lig.22).
Lig.22 (5 colonies), Marker 1Kb
  • Gel results: we chose 1st colony to grow a 9 ml overnight culture.

08/28/08

  • Glycerol stocks/miniprep for Lig.22.
  • We sent purified plasmids to Primm for sequencing.
  • We infected 9 ml of LB + Amp with 30 µl of R0062 glycerol stock.

08/29/08

  • Glycerol stock/miniprep for R0062.
Retrieved from "http://2008.igem.org/Team:UNIPV-Pavia/Notebook/Week15"