Team:Lethbridge/Notebook/Lab Work/Group3

From 2011.igem.org

(Difference between revisions)
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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
 +
 +
==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
 +
k331033-k331035 in pSB1C3
 +
P0440-K331033 in pSB1A2
 +
J04500-LUMAZINE SYNTHASE-DT in pSB1AK3
 +
pBAD-P0440 in pSB1A2
 +
LUMAZINE SYNTASE-DT-pBAD in pSB1A2
 +
 +
==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:
 +
K331035
 +
K331033
 +
Lumazine synthase - Dt
 +
P0440
 +
Control
 +
 +
EcoRI Restriction of:
 +
J04500-Agn 43 sample 1
 +
J04500 -Agn 43 Sample 2
 +
J04500 - Lumazine synthase - Dt sample 3
 +
K346007
 +
Lumazine synthase - Dt
 +
 +
Ran Restriction on gel to confirm sizes
 +
 +
Notebook team 3 insert picture Page 37

Revision as of 02:50, 15 September 2011



Contents

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
Lumazine - dt construct successfully inserted into pSB1C3 Glycerol stocked.

Add Gel Picture

July 8th

Performed antibiotic assay on lumazine - dt in pSB1C3
Samples from one colony grew in Kanamicyn; this stock was discarded

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 k331033-k331035 in pSB1C3 P0440-K331033 in pSB1A2 J04500-LUMAZINE SYNTHASE-DT in pSB1AK3 pBAD-P0440 in pSB1A2 LUMAZINE SYNTASE-DT-pBAD in pSB1A2

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: K331035 K331033 Lumazine synthase - Dt P0440 Control

EcoRI Restriction of: J04500-Agn 43 sample 1 J04500 -Agn 43 Sample 2 J04500 - Lumazine synthase - Dt sample 3 K346007 Lumazine synthase - Dt

Ran Restriction on gel to confirm sizes

Notebook team 3 insert picture Page 37
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