Difference between revisions of "IGEM:Harvard/2007/Laboratory Notebooks/Two Component System"

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6/27/07 - Ordered oligos to create a FecA promoter BioBrick that can be used to recombine with GFP to create a reporter system.
 
6/27/07 - Ordered oligos to create a FecA promoter BioBrick that can be used to recombine with GFP to create a reporter system.
  
The oligos ordered were as follows: Construct 1 5’- GTTTCTTCGAATTCGCGGCCGCTTCTAGAGatttcaccactgtaaggaaaataattcttatttc – 3’
+
The oligos ordered were as follows:  
 +
 
 +
Construct 1 5’- GTTTCTTCGAATTCGCGGCCGCTTCTAGAGatttcaccactgtaaggaaaataattcttatttc – 3’
  
 
Construct 2 5’- GTTTCTTCCTGCAGCGGCCGCTACTAGTAAGGGTAAAAAGGACAATCGAAATAAGAATTATTT – 3’
 
Construct 2 5’- GTTTCTTCCTGCAGCGGCCGCTACTAGTAAGGGTAAAAAGGACAATCGAAATAAGAATTATTT – 3’
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SV001 - FecA plus VF2
 
SV001 - FecA plus VF2
 +
 
SV002 - FecA plus VR
 
SV002 - FecA plus VR
 +
 
SV003 - FecR plus VF2
 
SV003 - FecR plus VF2
 +
 
SV004 - FecR plus VR
 
SV004 - FecR plus VR
 +
 
SV005 - FecI plus VF2
 
SV005 - FecI plus VF2
 +
 
SV006 - FecI plus VR
 
SV006 - FecI plus VR
 +
  
 
Insert site directed mutagenesis stuff here
 
Insert site directed mutagenesis stuff here
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7/9/07<br>
 
7/9/07<br>
 
-miniprep of site directed mutagenesis using QiaGen Miniprep kit
 
-miniprep of site directed mutagenesis using QiaGen Miniprep kit
 +
 
-purification of construct C1+C2 extension rxn R1 using QiaGen PCR purification protocol--labeled product 'FecA promoter biobrick'
 
-purification of construct C1+C2 extension rxn R1 using QiaGen PCR purification protocol--labeled product 'FecA promoter biobrick'
 +
 
-received E Coli AA93, plasmid pLCIRA, pGFPA' from Germany, waiting on instructions from Dr. Braun on how to plate them.  
 
-received E Coli AA93, plasmid pLCIRA, pGFPA' from Germany, waiting on instructions from Dr. Braun on how to plate them.  
 +
 
-grow E0240 in liquid culture
 
-grow E0240 in liquid culture

Revision as of 13:04, 9 July 2007

6/27/07 - Ordered oligos to create a FecA promoter BioBrick that can be used to recombine with GFP to create a reporter system.

The oligos ordered were as follows:

Construct 1 5’- GTTTCTTCGAATTCGCGGCCGCTTCTAGAGatttcaccactgtaaggaaaataattcttatttc – 3’

Construct 2 5’- GTTTCTTCCTGCAGCGGCCGCTACTAGTAAGGGTAAAAAGGACAATCGAAATAAGAATTATTT – 3’


6/27/07 - Grew bacteria (FecA, FecI, FecR) in liquid culture to prepare for sequencing tomorrow, inoculation in 2 ml LB and 2 ul amp 50mg/ml 1000x. [edit]


6/28/07 - Miniprepped to prepare for sequencing, nanodropped to confirm presence of DNA, sequencing rxns:

SV001 - FecA plus VF2

SV002 - FecA plus VR

SV003 - FecR plus VF2

SV004 - FecR plus VR

SV005 - FecI plus VF2

SV006 - FecI plus VR


Insert site directed mutagenesis stuff here

7/9/07
-miniprep of site directed mutagenesis using QiaGen Miniprep kit

-purification of construct C1+C2 extension rxn R1 using QiaGen PCR purification protocol--labeled product 'FecA promoter biobrick'

-received E Coli AA93, plasmid pLCIRA, pGFPA' from Germany, waiting on instructions from Dr. Braun on how to plate them.

-grow E0240 in liquid culture