Team:Hawaii/Notebook/2008-07-10

= Things we did today =

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 &mu;l 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 &mu;l 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"