Latest revision as of 14:50, 29 August 2015

Ferment It Yourself

Vitamin A	Vitamin B2	Vitamin B12
Phytase	Riboswitch	Differentiation on E. coli
Differentiation on S. cerevisiae	Manufacturing	Idli and Micro-organisms

Gel-Purification protocol with the QIAquick® Gel Extraction Kit

Excise the DNA band from the gel as precisely as possible and put it in a micro-centrifuge tube.
Add 3 volume of QG buffer (100mg of gel ~ 100ul) in the tube and incubate for 10 min at 50°C (or 10 min on a vortex).
Add one volume of isopropanol and mix.
Centrifugate the tube for 2 min at 14 000 rpm.
Carefully pipette the supernanant in a QIAquick column.
Put the column in a 2ml microcentrifuge tube and centrifuge it for 1 min at 14 000 rpm.
Discard the flow-through.
Add 750 uL of PE buffer in the column and centrifuge for 1 min and discard flow-through.
Centrifuge again for 2 min and put the column in a new 1.5uL microcentrifuge tube.
Elute the DNA by putting 50uL of water in the middle of the column and let it stand for 2 min.
Centrifugate for 2min at 10 000 rpm

@@ Line 10: / Line 10: @@
        <li>Add 3 volume of QG buffer (100mg of gel ~ 100ul) in the tube and incubate for 10 min at 50°C (or 10 min on a vortex).
        <li>Add one volume of isopropanol and mix.
-       <li>centrifugate the tube for 2 min at 14 000 rpm.
+       <li>Centrifugate the tube for 2 min at 14 000 rpm.
        <li>Carefully pipette the supernanant in a QIAquick column.
        <li>Put the column in a 2ml microcentrifuge tube and centrifuge it for 1 min at 14 000 rpm.
-       <li>Throw the supernatant.
+       <li>Discard the flow-through.
        <li>Add 750 uL of PE buffer in the column and centrifuge for 1 min and discard flow-through.
        <li>Centrifuge again for 2 min and put the column in a new 1.5uL microcentrifuge tube.