IGEM:Paris Liliane Bettencourt/Notebook/Paris 2010/2010/07/28: Difference between revisions
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<font color=red>Antoine</font> | |||
<br><u>Minipreps</u> Vf=35 uL | |||
*B0015 115.4 ng/uL | |||
*B0015 200.4 ng/uL | |||
*S200 174 ng/uL | |||
<br><u>Restriction digest 2h 37°C follow by inactivation at 80°C 20 min</u> Vf=20 uL | |||
{| | |||
|Sample name||DNA (1ug)||enzyme||Buffer||H2O||BSA | |||
|- | |||
|B0015 (200.4 ng/uL) || 5 ||1 X/P ||2(B3)|| 10.8 ||0.2 | |||
|- | |||
|B0015 || 5 ||1 X/P ||2(B3)|| 10.8 ||0.2 | |||
|- | |||
|B0015 || 5 ||1 X/P ||2(B3)|| 10.8 ||0.2 | |||
|- | |||
|B0015 || 5 ||1 X/P ||2(B3)|| 10.8 ||0.2 | |||
|- | |||
|S200 || 5.75||1 S/P ||2 (B2)||10.05||0.2 | |||
|- | |||
|S200 || 5.75||1 S/P ||2 (B2)||10.05||0.2 | |||
|} | |||
<br><u>Electrophoresis </u> | |||
<br><u>Gel purification </u> | |||
<br><u>Ligation follow by inactivation 65°C 10 min</u> Vf=10uL | |||
{| | |||
|Vecteur (10ng)||Insert(30ng)||Buffer ligase||H2O||T4 ligase | |||
|- | |||
|P1003(cut S+P) 0.7 || BOO15(cut X+P) 2.1 ||1 ||2(B3)|| 5.7||0.5 | |||
|- | |||
|C0012(cut S+P) 0.9 || BOO15(cut X+P) 2.1 ||1 ||2(B3)|| 5.5||0.5 | |||
|- | |||
|S173(cut S+P) 0.6 || BOO15(cut X+P) 2.1 ||1 ||2(B3)|| 5.8||0.5 | |||
|- | |||
|S200(cut S+P) 1.5 || BOO15(cut X+P) 2.1 ||1 ||2(B3)|| 4.9||0.5 | |||
|} | |||
<br><u>Transformation </u> | |||
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Revision as of 07:00, 5 August 2010
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Antoine
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