Team:Hawaii/Notebook/2008-10-13

From 2008.igem.org

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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. Ten microliters of PCR product were loaded into each lane.
  • Colony PCR of restreak plates
  • Bands ~600bp. Plasmid did not cut.
File:101308REdigest.jpg
EtBr stained 2% agarose gel ran at 60V for 2 hours. Thirty microliters of RE digested plasmid were loaded into each lane.
Krystle
  • RE digest of plasmid w/ EcoRI and PstI
  • RE digest of prpgt, rgt1 plasmids with EcoRI and PstI
  • Ran RE digests on gel
  • Bands did not correspond to what was supposedly loaded
Grace
  • PCR of prpgt #5, 7, 11, nrsg #6, rgt #1
  • Overnight RE digest of devices with EcoRI and PstI and rgt#1 with XbaI and PstI

Triparental Conjugation

Triparental conjugation plates. RP1, BB-pRL harboring E. coli and Synechocystis sp. PCC 6803 plated on 5% LB + BG-11 and incubated for 60 hours at 37C with 2% CO2
Grace
  • Restreaked 5%LB+BG-11 plates onto BG-11+sp2.5+sm2.5 plates for isolation of cyanos
  • Inoculated BG-11+sp2.5+sm2.5 liquid media with conjugated E. coli/cyanos
  • Incubated at 37C with 2% CO2 for 11 days. Liquid cultures were shaken at 100 rpm also.

Discussion

Quote of the Day

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


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