Revision as of 07:55, 11 July 2008

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Things we did today

Krystle

Grew up single colonies of Biobrick harboring E. coli for cryostocking and plasmid prep tomorrow.

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.

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.

we found this poured in the gel room today

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 ]

@@ Line 3: / Line 3: @@
 = Things we did today =
 == Wetlab work ==
-===Name of Task===
+===Growing up ''E. coli''===
-:<strong> name of person/people who performed the task</strong>
+:<strong> Krystle</strong>
-:* Summary of task and what was done. Link to experiment for detailed notes if necessary.
+:* Grew up single colonies of Biobrick harboring ''E. coli'' for cryostocking and plasmid prep tomorrow.
-:* e.g. worked on &lt;blah experiment link&gt;, PCR, ran gel
+===PCR===
+:<strong> Grace and Krystle</strong>
+:* 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===
+:<strong> Grace</strong>
+:* 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 [[Team:Hawaii/Initial_Synth._Oligo_Assembly|experiment]].
+:* Ran 0.8% gel with 20 &mu;l RE digested BBa_C0012 products.
+:* Excised bands from gels for DNA purification/extraction tomorrow.
 = Discussion =