Team:Paris Bettencourt/Notebook/Trojan Horse/Saturday 17th August.html

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Trojan Horse

17th August


Results

  • Transformation worked but no clone with the ligation product =>restart from the beginning, problem must be with one of the digested products.

  • Phage infectiveness : Experiment worked => results here

Repcr inserts / redigest vector

PCR KanR out of pCOLA (pT003)

5*50ul

34 H2O

10 Buffer 5x

1 dNTP

2.5 FW iT003

2.5 RV iT004

0.2 pCOLA

0.5 phusion

50ul Tot

Tm = 54°C

te = 45sec

PCR LacZ out of pUC18 (pT007)

5*50ul

34 H2O

10 Buffer 5x

1 dNTP

2.5 FW iT001

2.5 RV iT002

0.2 pUC18

0.5 phusion

50ul Tot

Tm = 54°C

te = 45sec

Gel of PCR Kan and LacZ

5 puits LacZ / 100bP+ / 5 puits Kan

Preparaiton of the vectors to clone the sRNA

PCR litmus (pT005)

5*50ul

40 H2O

10 Buffer 5x

1 dNTP

0.5 FW iT003

0.5 RV iT004

0.2 pCOLA (pT007)

0.5 phusion

50ul Tot

Tm = 64°C

te = 2min10

Results

  • Transformation worked but no clone with the ligation product =>restart from the beginning, problem must be with one of the digested products.

  • Phage infectiveness : Experiment worked => results here

Repcr inserts / redigest vector

PCR KanR out of pCOLA (pT003)

5*50ul

34 H2O

10 Buffer 5x

1 dNTP

2.5 FW iT003

2.5 RV iT004

0.2 pCOLA

0.5 phusion

50ul Tot

Tm = 54°C

te = 45sec

PCR LacZ out of pUC18 (pT007)

5*50ul

34 H2O

10 Buffer 5x

1 dNTP

2.5 FW iT001

2.5 RV iT002

0.2 pUC18

0.5 phusion

50ul Tot

Tm = 54°C

te = 45sec

Gel of PCR Kan and LacZ

5 puits LacZ / 100bP+ / 5 puits Kan

-->img 622<--

Preparaiton of the vectors to clone the sRNA

PCR litmus (pT005)

5*50ul

40 H2O

10 Buffer 5x

1 dNTP

0.5 FW iT003

0.5 RV iT004

0.2 pCOLA (pT007)

0.5 phusion

50ul Tot

Tm = 64°C

te = 2min10