Team:Hawaii/Notebook/2008-08-17

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Things we did today

Wetlab work

Verification of transformants

Grace
EtBr stained 2% agarose gel ran at 95V for 1 hour. Five microliters of PCR reaction were loaded into each well.
DNA Plate Total colony forming units Blue colonies
BB-pRL1383a + J33207 LB + sp100 + sm50 + IPTG (1mM) + X-gal (2mM) 105 0
BB-pRL1383a + J33207 LB + sp100 + X-gal (2mM) 44 0
BB-pRL1383a (negative control) LB + sp100 + sm50 + IPTG (1mM) + X-gal (2mM) 44 0
no DNA (negative control) LB + sp100 + sm50 + IPTG (1mM) + X-gal (2mM) 0 0
  • Colony PCR to verify insert
  • ~325bp band is due to pRL1383a plasmid
  • BB-pRL1383a + J33207 bands are smaller than (-) control bands, potential success in replacing GFP w/ lac device
  • (-) bands due to contamination (no template added to PCR rxn) and GFP (religated plasmid, one RE did not cut)
  • Restreaked colonies to purify
  • Grew up colonies on TB+sp100 for plasmid prep

Discussion

Quote of the Day

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson


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