Team:Hawaii/Notebook/2008-08-17

From 2008.igem.org

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Things we did today

Wetlab work

Verification of transformants

Grace
EtBr stained 2.5% agarose gel ran at 95V for 1 .5 hours. 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
  • X-gal did not work?
  • Colony PCR to verify insert
  • ~320bp 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
  • Restreaked colonies to purify
  • Grew up colonies in TB+sp100 liquid media for plasmid prep on Tuesday

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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