IGEM:Peking/2007/Switch-Notebook/2007-7-24

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Experimenter: LD & CXY

Contents

Contents

Precipitate to retract: pLX007/8/9^^(XhoI/SalI), lacZa^^(XhoI/SalI)

Overnight ligation of pLX007/8/9^^ and lacZa^^

  • Plasmid: 4.5uL, lacZa: 0.5uL

Retract from electrophoresis jel: lacZa-T^(XhoI/SalI)

Image:PKU Switch 07-07-24 LacZa retract.jpg

Not retracted because the concentration is too low.

Temperature PCR of RS1-3, RD1-3 from Prec-T

  • Template: 1uL
  • Buffer: 5uL
  • dNTP: 5uL
  • Primer_F Rec_S_1_F or Rec_D_1_F: 1uL
  • Primer_R Rec_S_1_Rnew: 1uL
  • Ex-taq: 0.25uL
  • H2O: 37uL
  • Total: 50.25uL
  • Product: RS1, RD1
  • Temperature gradient: 53.2C, 58.1C, 63.5C, 68.1C

Image:PKU Switch 07-07-24 RD1-4 SD1-4 PCR.jpg

l to r: RD1(1-4), RS1(1-4), 100kb ladder

retract: RS1(53.2C), RD1(53.2C)

  • Template: 1uL
  • Buffer: 5uL
  • dNTP: 5uL
  • Primer_F Rec_S_2/3_F or Rec_D_2/3_F: 1uL
  • Primer_R Rec_S_2/3_Rnew: 1uL
  • Ex-taq: 0.25uL
  • H2O: 37uL
  • Total: 50.25uL
  • Product: RS2/3, RD2/3
  • Temperature gradient: 68C

Image:PKU Switch 07-07-24 RS2RS3RD2RD3.jpg

l to r: RS2, RS3, RD2, RD3, 100kb ladder

retract: RS2, RD2

Overnight ligation of RS1, RD1, RS2, RD2 with pMD18-T

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