Team:UNIPV-Pavia/Notebook/Week16

From 2008.igem.org

(Difference between revisions)
 
(One intermediate revision not shown)
Line 46: Line 46:
!|[[Team:UNIPV-Pavia/Notebook/Week20|Week 20]]
!|[[Team:UNIPV-Pavia/Notebook/Week20|Week 20]]
!|[[Team:UNIPV-Pavia/Notebook/Week21|Week 21]]
!|[[Team:UNIPV-Pavia/Notebook/Week21|Week 21]]
 +
|-
 +
!|[[Team:UNIPV-Pavia/Notebook/Week22|Week 22]]
 +
!|[[Team:UNIPV-Pavia/Notebook/Week23|Week 23]]
 +
!|[[Team:UNIPV-Pavia/Notebook/Week24|Week 24]]
|}
|}
Line 57: Line 61:
**Lig.22-R0062
**Lig.22-R0062
**Lig.31-Lig.22
**Lig.31-Lig.22
-
*but we didn't have enough DNA for 22 and 31 plasmids...
+
*but we didn't have enough Lig.22 and Lig.31 plasmids...
*We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.
*We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.

Latest revision as of 21:27, 26 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 22 Week 23 Week 24



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 Lig.22 and Lig.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.