Team:Lethbridge/Notebook/Lab Work/Group3 - 2011.igem.org
Team:Lethbridge/Notebook/Lab Work/Group3
From 2011.igem.org
(Difference between revisions)
Line 77:
Line 77: [[Team:Lethbridge/Notebook|PCR]] <br />
[[Team:Lethbridge/Notebook|PCR]] <br />
- PCR'd out lumazine - dt <br />
+ * PCR'd out lumazine - dt <br />
- Used Prefix and antisense suffix
+ * Used Prefix and antisense suffix
Ran samples on Gel to confirm sizes
Ran samples on Gel to confirm sizes
Line 90:
Line 90: ==July 25th, 2011==
==July 25th, 2011==
- ran 4 samples on an agarose gel
+ Ran 4 samples on an agarose gel: <br />
+ D22 <br />
+ T22 <br />
+ D24 <br />
+ T24 <br />
- Preformed a restriction with EcoRI and PstI for Ryan
+ Preformed a restriction with EcoRI and PstI for Ryan <br />
+ Ran on 1% agarose gel
==July 28th, 2011==
==July 28th, 2011==
Line 124:
Line 129: Notebook team 3 insert picture Page 37
Notebook team 3 insert picture Page 37
+
+ ==August 2nd, 2011==
+
+ Transfer of enhanced lumazine synthase from pET-28a into pSB1C3
+
+ [[Team:Lethbridge/Notebook|Restricted]] with EcoR1 and Pst1
+
+ Ran on 1% TAE agarose gel for 90 minutes at 80V
Revision as of 03:03, 15 September 2011
June
June 21st, 2011
Restriction, Ligation, and Transformation
Lumazine synthase - double terminator and pBAD - RBS into pSB1C3
1. Restriction
Restricted lumazine synthase - dt out of pSB1K3 and pBAD - RBS out of pSB1T3 with EcoR1 and Pst1
Restricted destination plasmid pSB1C3 with EcoR1 and Pst1
2. Ligation
Ligated lumazine synthase-dt into destination plasmid pSB1C3
Ligated pBAD-RBS into destination plasmid pSB1C3
3. Transformation
Transformed lumazine synthase - dt in pSB1C3 into DH5∝
Transformed pBAD-RBS in pSB1C3 into DH5∝
June 24th, 2011
Picked cells from June 21st transformation
June 25th, 2011
Minipreped Cells picked on June 25th, 2011
June 28th
Restriction digest and ran on gel
Add Gel Picture
July 8th
Performed antibiotic assay on lumazine - dt in pSB1C3
PCR
PCR'd out lumazine - dt Used Prefix and antisense suffix
Ran samples on Gel to confirm sizes
July 10th, 2011
PCR'd out Lumazine Synthase - Dt and RFP
Ran on a 1% agarose gel for 45min at 70volts
July 25th, 2011
Ran 4 samples on an agarose gel:
Preformed a restriction with EcoRI and PstI for Ryan
July 28th, 2011
picked cells from plates and ran a Colony PCR of:
July 29th, 2011
Ran PCR samples from July 28th on a 1X TBE Agarose gel
For control comparision of parts from July 28th PCR we ran a PCR of:
EcoRI Restriction of:
Ran Restriction on gel to confirm sizes
Notebook team 3 insert picture Page 37
August 2nd, 2011
Transfer of enhanced lumazine synthase from pET-28a into pSB1C3
Restricted with EcoR1 and Pst1
Ran on 1% TAE agarose gel for 90 minutes at 80V
"
