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Team:Hawaii/Notebook/2008-07-10
From 2008.igem.org
| Projects | Events | Resources | ||
|---|---|---|---|---|
| Sponsors | Experiments | Milestones | Protocols | |
| Notebook (t) | Meetings (t) |
Things we did today
Wetlab work
Growing up E. coli
- Krystle
- Grew up single colonies of Biobrick harboring E. coli for cryostocking and plasmid prep tomorrow.
PCR
- Grace and Krystle
- Ran 10 μl PCR reactions with Green Taq for GFP site directed mutagenesis and priming out the Biobrick sites from BBa_C0012 (for construction of our own Biobrick vector)
- Forgot to keep everything on ice. May have non-specific amplifications. Will run on a gel tomorrow to check.
Redid ligation/restriction digest of annealed products
- Grace
- Ligated 1:1 and 1:10 dilutions of annealed product (20-30 min ligation) then restriction digested w/ XbaI and PstI using full RE buffer amounts recommended by NEB (2.5 hours digest).
- Heated digested products in 95C water bath for 10 min before running on gel.
- Ran 3% gel with RE products (both ligation dilutions) and annealed but not ligated products. Gel still had ladders. See experiment.
- Ran 0.8% gel with 20 μl RE digested BBa_C0012 products.
- Excised bands from gels for DNA purification/extraction tomorrow.
Discussion
Quote of the Day
Looks like someone is developing a comb-less gel electrophoresis protocol. - NW, KLS
[http://manoa.hawaii.edu/ 
][http://manoa.hawaii.edu/ovcrge/ 
][http://www.ctahr.hawaii.edu 
]
