Ferment It Yourself

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

Yeast DNA extraction using DNeasy® Blood & Tissue Kit

Centrifuge a maximum of 5x10⁶ cells for 5 minutes at 300g (190rpm). Resuspend in 200µL PBS. Add 20µL zymolyase.
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.
vortex 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.
Throw the supernatant.
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

Team:Paris Bettencourt/Protocols/dneasy-blood-and-tissue-kit