Team:Hawaii/Notebook/2008-07-15

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Things we did today

Wetlab work

Oligonucleotide experiment, continued

Grace
  • Ran overnight RE digests of pRL1383a and BBa_C0012 on EtBr stained 2% agarose gel at 95V for 1 hour
  • BBa_C0012 vector okay
  • pRL1383a vector band ~9kb; no band observed at ~350bp. RE did not cut?
  • Gel purified pRl1383a, BBa_C0012, GFP fusion, and Biobrick segments
  • Used nanodrop spectrometer to determine DNA concentration (below)
  • Ligated 7 μl Biobrick segment with 0.5 μl pRL1383a and 7 μl GFP fusion with 2 μl BBa_C0012 vector in 20 μl ligation reactions
  • Transformed DH5α using 10 μl ligation reactions
  • Plated GFP/C0012 vector on LB + amp100
  • Plated BB segment/pRL1383a on LB + sp100
  • Counted colonies from yesterday's pnir, slr2016, pilA assembly transformation (see experiment)
  • Colony PCR'd colonies to verify presence of desired products

Transformation

Margaret

  • Transformation of DB3.1 competent cells pSB3K3, pSB1A2, pSB1AK3, (+)pUC18, (-) no plasmid, because transformation from 7/14/08 yielded only 1 colony from pSB3K3, nothing from pSB1A2 & pSB1AK3.

Plasmid prep

Krystle

  • Resuspended yesterday's plasmid prep in 100 μl TE and stored in -20C
  • Ran plasmid preps on gel to verify
  • Grew up pnir, slr2016, and pilA transformants in LB in preparation for plasmid prep tomorrow

Restriction Digest

Krystle

  • RE digested BBa_B0034, BBa_B0024, and BBa_E0040 with XbaI overnight

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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