From 2008.igem.org

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Things we did today

Wetlab work

Verification of transformants

EtBr stained 2% agarose gel ran at 95V for 1.5 hours. Five microliters of PCR reaction were loaded into each lane.

Grace

• Colony PCR of transformants
• Ran PCR reaction on 2% agarose gel

• Can't tell 15bp difference between GFP and rbs+GFP

• Restreaked single colony and inoculated for plasmid prep to confirm

• nir+rbs colonies #11-19 appear promising (larger than nir)

• Restreaked colony #19 and inoculated for plasmid prep to confirm

Plasmid prep of J33207

Grace

Overnight RE digest

Grace

• BB-pRL1383a and J33207 with XbaI and PstI
• pSB1A2 with EcoRI and PstI
• GFP, GFPf, nir with EcoRI and SpeI

Made LB+amp₁₀₀ plates

Grace

Ligation

Margaret

• ligation of rep+pSB1A3, P1lytic region+pSB1A3, aada(BB&pRL1383a)+pSB1A3

omega interposon

• streak on Sm&Sp plate to confirm insertion.

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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