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Things we did today

Wetlab work

Plasmid prep

Krystle

• Finished plasmid prep of BB-pRL1383a, nir, GFPf from yesterday

Cyanobacteria seed stocks

Krystle

• Reinoculated liquid and solid media seed stocks

Verification of transformants

EtBr stained 2.5% agarose gel ran at 95V for 75 min. Ten microliters of the PCR reaction were loaded into each well.

Grace

• Colony PCR of transformants
• Ran on EtBr stained 2.5% agarose gel at 95V for 75 min.

• Looks like nir+rbs and I14032+rbs worked. YAY :o). Will prep for sequencing tomorrow.
• GFP, GFPf, J33207 don't look like they went in. Will sequence to confirm anyway.

Plasmid Prep

Margaret

Plasmid Prep of pSMC121.

• Re-did yesterday's plasmid prep of pSMC121 because I don't believe the band was big enough. I e-mailed Dr. Callahan to get the exact size, but I know, because it contains the omega interposon, that it is at least 2kb.

Extraction from Gel

Margaret

• extracted 3 bands of 50ul reaction for rep and oriV first by cutting out using long wave UV for visualization.
• Purification from gel with Qiagen Minelute gel purification kit.

PCR

Large scale PCR for amplification of P1 lytic and aadA region.

Margaret

• Large-scale PCR amplification of aadA (it was lost in a gel cutting mishap) and p1 lytic.

Discussion

Quote of the Day

Gernot: "So would you eat the E. coli to save them if we lose the -80C in like a war?"
Loren: "Well, I'll eat them, but you'll be collecting them."

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

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