User:Michael Verhoeven/Notebook/iGEM 2009 M. Verhoeven Parts/2009/07/06

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Transformation of E. coli TOP10 with BBa_J23109, BBa_J23100 and BBa_J23106

  • BBa_J23109 iGEM plate 2, 2G
  • BBa_J23100 iGEM plate 1, 18C
  • BBa_J23106 iGEM plate 1, 18O

All dry plasmid DNA in the wells was dissolved in 15ul sterile diH2O, and transfered to 500ul cups.

  • Four cups with 50ul competent E.coli TOP10 cells were thawed on ice, and 2ul of dissolved plasmid was added.
  • After 30 min. on ice, the cups were put in a heat block at 37C for 5 min. and followed by 5 min. on ice.
  • To the cells, 800ul TY medium was added and incubated at 37C for 1 hour.
  • The cells were centrifuged for 1 min. at full speed and the cells were resuspended in 200μL TY-medium, and 100μL was plated on TY-Agar-Amp plates. The plates were stored at 37C o.n.
  • The remaining 100ul was diluted 100x and 100μL was plated on TY-Agar-Amp plates and incubated o.n. at 37C.

Inoculation of TY-medium for Glycerol Stocks

  • GVP cluster vector BBa_J61035 (2x) in E.coli TOP10
  • Terminator vector ? (2x) in E.coli TOP10
  • 4.5 ml TY-medium with Amp. was inoculated with a colony grown on TY-Agar-Amp. plates o.n.
  • The tubes were put in a waterbath at 37C (150 rpm) and grown o.n.