IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/12: Difference between revisions
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* We removed our cultures of e. coli containing Miraculin/Brazzein C/N YFP and NosT & Stop and e. coli containing Miraculin/Brazzein and StrepII tag from the 37°C shaker. | * We removed our cultures of e. coli containing Miraculin/Brazzein C/N YFP and NosT & Stop and e. coli containing Miraculin/Brazzein and StrepII tag from the 37°C shaker. | ||
* Glycerol stocks were made (666 μL and 333 μL) | * Glycerol stocks were made (666 μL glycerol and 333 μL cells) | ||
===Miniprep=== | ===Miniprep=== | ||
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Brazzein C StrepII 1: 209.3 ng/μL | Brazzein C StrepII 1: 209.3 ng/μL | ||
Brazzein C StrepII 2: 346.4 ng/μL | Brazzein C StrepII 2: 346.4 ng/μL | ||
===Digestion=== | |||
Revision as of 07:47, 13 July 2010
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Team AllergyToday, we received sequences for the allergen , intron, and amiRNA inserts. Since 4 of the 8 allergen inserts were not correctly done (and were not set for sequencing) and we want to repeat the inserts that DID work, we will redo ligations for the entire panel. Procedures
ResultsSequencing Results
Full report (annotated sequences): File:Report.zip Redo Ligations w/ phosphatized backbone for entire allergen panel, PME did not grow, we will not use it since we have good PDK and PAL introns. PAL and PDK will be digested and inserted into a plasmid containing a sense strand of an allergen. Add PDK to amiRNA vectors if sequences match up
Team Flavor
MiniprepODs Miraculin C YFP and NosT & Stop 1: 414.4 ng/μL Miraculin C YFP and NosT & Stop 2: 304.0 ng/μL Miraculin N YFP and NosT & Stop 1: 428.6 ng/μL Miraculin N YFP and NosT & Stop 2: 421.0 ng/μL Brazzein C YFP and NosT & Stop 1: 572.8 ng/μL Brazzein C YFP and NosT & Stop 2: 580.3 ng/μL Brazzein N YFP and NosT & Stop 1: 348.9 ng/μL Brazzein N YFP and NosT & Stop 2: 639.6 ng/μL Miraculin N StrepII 1: 888.6 ng/μL Miraculin N StrepII 2: 614.4 ng/μL Miraculin C StrepII 1: 677.1 ng/μL Miraculin C StrepII 2: 222.1 ng/μL Brazzein N StrepII 1: 352.6 ng/μL Brazzein N StrepII 2: 331.6 ng/μL Brazzein C StrepII 1: 209.3 ng/μL Brazzein C StrepII 2: 346.4 ng/μL
Digestion |