Team:HokkaidoU Japan/Notebook/September8

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Colony PCR

Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids

  • September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version)
  • 1-3A
  • Retried September 6th transformation
  • pUC119

Confirmation of pSB1C3

Because pSB1C3 PCRed from 1-3A(RFP reporter) was used there was posibility of contamination by template which would also produce red colonies.

  • H buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
  • M buffer treated 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
  • H buffer treated 2 uL → Transformation
  • H buffer treated 2 uL → Transformation

PCR of GFP

  • GFP reporter BBa_I13522 pSB1A2 2-8A 937bp
  • GFP protein BBa_E0840 pSB1A2 1-12O 878bp
PCR cocktail was same as September 1st's