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Team:Paris Bettencourt/Notebook/Trojan Horse/Saturday 17th August.html
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5 puits LacZ / 100bP+ / 5 puits Kan | 5 puits LacZ / 100bP+ / 5 puits Kan | ||
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<strong>Preparaiton of the vectors to clone the sRNA</strong> | <strong>Preparaiton of the vectors to clone the sRNA</strong> | ||
Latest revision as of 20:17, 3 October 2013
Trojan Horse
ASDF17th 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
