Latest revision as of 00:57, 30 October 2008

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Things we did today

EtBr stained 2% agarose gel ran at 60V for 2 hours. Thirty microliters of RE digested DNA were loaded into each well.

Grace

Krystle

Grace

prpgt, nrsg, sgt, and pgt sequences all returned inserts of E. coli genomic DNA instead of desired construct

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson

[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]

@@ Line 4: / Line 4: @@
 == Wetlab work ==
 ===Construction of secretion device (cont.)===
-:<strong> Grace</strong>
 [[Image:101408REdigest.png|right|thumb|300px|EtBr stained 2% agarose gel ran at 60V for 2 hours. Thirty microliters of RE digested DNA were loaded into each well.]]
+:<strong> Grace</strong>
 :* Ran RE digests on gel
 :* Extracted rgt1 and prpgt5 from gel
@@ Line 14: / Line 15: @@
 ::* nrsg6 (from 10/11) + BBpRL1383a-1
 ::* J33207 + pRL1383a
-:* Transformed ''E. coli''
-::* Used 5 &mu;l ligation reaction (those containing BBpRL1383a-1) to transform DB3.1 cells
-::* Used 1 &mu;l ligation reaction (J33207 + pRL1383a) to transform DH5&alpha; cells
-:::* Plated 25 &mu;l, 50 &mu;l, 75 &mu;l, and 100 &mu;l of transformed cells on LB+sp<sub>2.5</sub>+sm<sub>2.5</sub> plates
+===Plasmid Prep===
+:<strong>Krystle</strong>
+:* Plasmid Prepped 2 cultures of BBpRL1383
+:* Nanodrop concentration is 330 ng/ul
+:* Gel verification for plasmid shows blank lanes
+==Drylab Work==
+===Sequencing analysis===
+:<strong>Grace</strong>
+:* prpgt, nrsg, sgt, and pgt sequences all returned inserts of ''E. coli'' genomic DNA instead of desired construct
 = Discussion =