From 2008.igem.org

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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 ]

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