IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/06/21: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 6: Line 6:
| colspan="2"|
| colspan="2"|
==Team Allergy==
==Team Allergy==
===Procedures===


PCR (see gel result below)
Today, the objective is to obtain PCR products of Arabadopsis allergens LTP 1, Ger 3, and Bet v1.
 
[[Image: Ltp bet ger 621.jpg|thumb|left| lanes 2,11,12 were successful]]
 
 
 
 
 
 
 
 
 


1. Obtain wildtype Arabadopsis genomic DNA
2. PCR for each LTP 1, Ger 3, and Bet v1




[[Image: Ltp bet ger 621.jpg|thumb|left| PCR of LTP, Ger 3, Bet v1 and v2, sense and antisense. Only lanes 2,11,12 were successful]]


==Team Genetic Fence==
==Team Genetic Fence==

Revision as of 11:30, 23 June 2010

iGEM iGarden <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Team Allergy

Procedures

Today, the objective is to obtain PCR products of Arabadopsis allergens LTP 1, Ger 3, and Bet v1.

1. Obtain wildtype Arabadopsis genomic DNA 2. PCR for each LTP 1, Ger 3, and Bet v1


PCR of LTP, Ger 3, Bet v1 and v2, sense and antisense. Only lanes 2,11,12 were successful

Team Genetic Fence

Barnase and Barstar

Barnase and Barstar Plasmids from ADDGENE arrived in mail

pMT316 - Barstar. Info from ADDGENE

pMT413 - Barnase, Barstar. Info from ADDGENE

pMT1002 - Barnase, Barstar. Info from ADDGENE


Notes from lab meeting

LVA tail for Cre


GAL4 DBD Innoculation

Pipetted 5mL of LB+Amp each into capped cc tubes, and mixed bacterial scrapings from 5 separate colonies of transformed GAL4 DNA binding domain E.Coli. Set to shake at 37°C overnight.


Cre, Lox66, Lox 71, GFP, and Luciferase Bacterial Transformation

Performed bacterial transformation according to Silver:_Bacterial_Transformation, however we used 1μL in step 3, and used entire tube of TOP10 competent cells instead of only 10-15μL.


Transformed the following each into its own tube of TOP10 E.Coli:

Cre DNA Recombinase

  • biobrick BBa_J61047, located on plate 1, well 5D, plasmid pSB1A2

Lox71

  • biobrick BBa_I718017, located on plate 1, well 17J, plasmid pSB1A2

Firefly Luciferase

  • biobrick BBa_I712019, located on plate 1, well 10H, plasmid pSB1AK8.

GFP mutant (phenotype identical to wildtype)

  • biobrick BBa_E0040, located on plate 1, well 14K, plasmid pSB1A2.

Lox66

  • biobrick BBa_I718016, located on plate 1, well 17H, plasmid pSB1A2.


Labeled each plate LB+Amp, its contents (ie Lox71 or Firefly Luciferase), MP, JW, OM, 6-21-10

Set all five plates to incubate at 37°C overnight.




Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:Harvard/2009/Notebook/Harvard_iGEM_2010/2010/06/21&oldid=426720"