Team:UNIPV-Pavia/Notebook/Week16

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<br>
<br>
*We had to perform these ligations:
*We had to perform these ligations:
-
**22-BBa_R0062
+
**Lig.22-R0062
-
**31-22
+
**Lig.31-Lig.22
*but we didn't have enough DNA for 22 and 31 plasmids...
*but we didn't have enough DNA for 22 and 31 plasmids...
-
*We infected 9 ml of LB + Amp with 30 µl of 22 and 31 glycerol stocks.
+
*We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.
'''09/2/08'''
'''09/2/08'''
<br>
<br>
-
*Glycerol stocks/miniprep for 22 and 31.
+
*Glycerol stocks/miniprep for Lig.22 and Lig.31.
*Plasmid digestion for:
*Plasmid digestion for:
-
**22 (E-S)
+
**Lig.22 (E-S)
-
**22 (X-P)
+
**Lig.22 (X-P)
-
**31 (S-P)
+
**Lig.31 (S-P)
-
**BBa_R0062 (E-X)
+
**R0062 (E-X)
*Run/gel extraction.
*Run/gel extraction.
*Ligations:
*Ligations:
-
**31-22 (="35")
+
**Lig.31-Lig.22 (=Lig.35)
-
**22-BBa_R0062 (="30")
+
**Lig.22-R0062 (=Lig.30)
-
*We incubated ligations at 16°C overnigth.
+
*We incubated ligations at 16°C overnight.
'''09/3/08'''
'''09/3/08'''
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*Gel results:
*Gel results:
-
**30 (3rd colony)
+
**Lig.30 (3rd colony)
-
**35 (gel showed an unexpected contaminant...we decided to re-perform this ligation)
+
**Lig.35 (gel showed an unexpected contaminant...we decided to re-perform this ligation)
'''09/5/08'''
'''09/5/08'''
<br>
<br>
-
*Glycerol stock/miniprep for 30.
+
*Glycerol stock/miniprep for Lig.30.
-
*We sent 30(3) to Primm for sequencing.
+
*We sent Lig.30 (3rd col) to Primm for sequencing.

Revision as of 13:20, 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:
  • 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.
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