Revision as of 00:38, 18 August 2008

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Things we did today

Grace

EtBr stained 2.5% agarose gel ran at 95V for 1 .5 hours. Five microliters of PCR reaction were loaded into each well.

DNA	Plate	Total colony forming units
BB-pRL1383a + J33207	LB + sp₁₀₀ + sm₅₀ + IPTG (1mM) + X-gal (2mM)	105
BB-pRL1383a + J33207	LB + sp₁₀₀ + X-gal (2mM)	44
BB-pRL1383a (negative control)	LB + sp₁₀₀ + sm₅₀ + IPTG (1mM) + X-gal (2mM)	44
no DNA (negative control)	LB + sp₁₀₀ + sm₅₀ + IPTG (1mM) + X-gal (2mM)	0

~320bp band is due to pRL1383a plasmid
BB-pRL1383a + J33207 bands are smaller than (-) control bands, potential success in replacing GFP w/ lac device

(-) bands due to contamination (no template added to PCR rxn) and GFP (religated plasmid, one RE did not cut)

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 5: / Line 5: @@
 ===Verification of transformants===
 :<strong> Grace</strong>
-[[Image:081708colonyPCR.jpg|right|thumb|250px|EtBr stained 2% agarose gel ran at 95V for 1 hour. Five microliters of PCR reaction were loaded into each well.]]
+[[Image:081708colonyPCR.jpg|right|thumb|250px|EtBr stained 2.5% agarose gel ran at 95V for 1 .5 hours. Five microliters of PCR reaction were loaded into each well.]]
 {| class=wikitable border=1 align=center
 ! DNA
@@ Line 33: / Line 33: @@
 |}
 :* Colony PCR to verify insert
-::* ~325bp band is due to pRL1383a plasmid
+::* ~320bp band is due to pRL1383a plasmid
 ::* BB-pRL1383a + J33207 bands are smaller than (-) control bands, potential success in replacing GFP w/ lac device
 :::* (-) bands due to contamination (no template added to PCR rxn) and GFP (religated plasmid, one RE did not cut)