Team:Paris Bettencourt/Protocols/PCR purification
Ferment It Yourself
iGEM Paris-Bettencourt 2O15
- Background
- Design
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Notebook
Vitamin A Vitamin B2 Vitamin B12 Phytase Riboswitch Differentiation on E. coli Differentiation on S. cerevisiae Manufacturing Idli and Micro-organisms -
PCR purification protocol
- Add 5 volumes of resuspension buffer to 1 volume of PCR product in an 1.5mL microcentrifuge tube, mix by pipetting up and down
- Transfer in a centrifugation column
- Centrifuge 1 min at 14000 rpm
- Throw the filtration product
- Add 700μL of washing solution
- Centrifuge 1 min at 14000 rpm
- Throw the filtration product
- Add 500μL of washing solution
- Centrifuge 1 min at 14000 rpm
- Throw the filtration product
- Centrifuge 1 min at 14000 rpm
- Throw the filtration product
- Put the column in a sterile 1.5mL microcentrifuge tube
- Add 45μL of DNAse/RNAse free water on the membrane
- Wait 2 minutes
- Centrifuge 2min at 10000rpm
- Discard the column, DNA is saved in water </td>