From 2008.igem.org

Latest revision as of 04:53, 18 July 2008

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Things we did today

Wetlab work

PCR

Grace

PCR products on an EtBr stained 4% agarose gel run at 119V for 27 min then 95V for 25 min. Ten microliters of PCR reactions were loaded into each well.

• Colony PCR for GFP fusion brick and new pRL1383a to verify inserts
• Sequencing (25 mu;l reactions)

• nir promoter
• slr2016
• pilA
• BBa_B0015
• BBa_B0024
• BBa_B0030
• BBa_C0012
• BBa_E0040
• BBa_J04430
• BBa_J33207
• BBa_R001

• Ran all PCR products on a 4% gel to check

• Gel ran funny -- loading dye ran 2x as fast as the DNA itself

• 119V too much to handle? Wavy bands

• Poor ladder resolution on gel
• No bands seen for GFP fusion, pRL1383a, R0010
• Incorrect bands for B0015 (multiple bands, what are the two bands at the top?) and J33207 (size too small, stuff still in wells)
• Will redo plasmid preps for R0010, B0015, J33207
• Will redo PCR reaction tomorrow for pRL1383a, GFP fusion, B0015, J33207, R0010

Plasmid Prep

Grace

• Inoculated 7 ml LB + amp₁₀₀ with single colony of B0015, R0010, J04430 for redo of plasmid prep tomorrow

Restreak to Purify

Grace

• Restreaked GFP fusion and pRL1383a (MCS replaced) transformants to purify

pSB3K3 plasmid prep

Margaret

Discussion

Quote of the Day

At the team meeting today:

"We have cookies, we can start."

Ten cookies later...

"The more cookies I have, the better I think!"

- Dr. Callahan

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

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