Latest revision as of 01:05, 30 October 2008

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Things we did today

Wetlab work

Insertion of lac-GFP device into BB-pRL1383a

EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Thirty microliters of the RE digest reaction were loaded into each well.

EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Ten microliters of PCR product were loaded.

Grace

Ran RE digests on gel

Bands were all incorrect size

Reinoculated TB+sp₁₀₀ for plasmid prep tomorrow
PCR of J04430 from filter paper to verify part; used H/B primers

Team:Hawaii/Antibiotic test for BB-pRL1383a

Grace

Made LB+sp_variable plates with 50 μl 20X X-gal
Transformed 100 μl DH5α with 2 μl BB-pRL1383a (4 ng)
Plated 50 μl cells + SOB on each LB+sp plate; plated 20 μl untransformed cells + SOB on each control plate
Incubated at 37C for 2 days

Construction of GFP secretion device

Krystle

Gel purified overnight restriction digested nir+rbs

Loaded all of restriction digest into a 2% gel, which appeared blank after running for 1.5 hours

Re-PCR nir+rbs and slr+gfpf from plasmid prep
Restriction digest

nir+rbs with EcoRI and SpeI
slr+gfpf with XbaI and PstI

Discussion

Quote of the Day

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 ==
 ===Insertion of lac-GFP device into BB-pRL1383a===
+[[Image:102108REdigests.png|right|thumb|150px|EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Thirty microliters of the RE digest reaction were loaded into each well.]][[Image:102108J04430.png|right|thumb|150px|EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Ten microliters of PCR product were loaded.]]
 :<strong>  Grace</strong>
-[[Image:102108REdigests.png|right|thumb|EtBr stained 0.8% agarose gel ran at 60V for 90 minutes. Thirty microliters of the RE digest reaction were loaded into each well.]]
 :*Ran RE digests on gel
-::* [[quantification of bands]]
+::*Bands were all incorrect size
-:* Extracted bands from gel
+:* Reinoculated TB+sp<sub>100</sub> for plasmid prep tomorrow
-:* Ligated J04430 and BB-pRL1383a
+:* PCR of J04430 from filter paper to verify part; used H/B primers
-:* Used 5&mu;l to transform DH5&alpha;
-===Antibiotic test for BB-pRL1383a===
+===[[Team:Hawaii/Antibiotic test for BB-pRL1383a]]===
 :<strong> Grace</strong>
-:* Made LB+sp<sub>variable</sub> plates with 40 &mu;l 20X X-gal
+:* Made LB+sp<sub>variable</sub> plates with 50 &mu;l 20X X-gal
-:* Transformed 100 &mu;l DH5&alpha; with 2 &mul; BB-pRL1383a [[ng]]
+:* Transformed 100 &mu;l DH5&alpha; with 2 &mu;l BB-pRL1383a (4 ng)
-:* Plated 50 &m;ul cells + SOB on each plate
+:* Plated 50 &mu;l cells + SOB on each LB+sp plate; plated 20 &mu;l untransformed cells + SOB on each control plate
+:* Incubated at 37C for 2 days
+===Construction of GFP secretion device===
+:<strong>Krystle</strong>
+:* Gel purified overnight restriction digested nir+rbs
+::* Loaded all of restriction digest into a 2% gel, which appeared blank after running for 1.5 hours
+:* Re-PCR nir+rbs and slr+gfpf from plasmid prep
+:* Restriction digest
+::*nir+rbs with EcoRI and SpeI
+::*slr+gfpf with XbaI and PstI
 = Discussion =

Team:Hawaii/Notebook/2008-10-21

From 2008.igem.org