Team:Paris Bettencourt/Protocols/titration-acid-phytic
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 Titration of phytc acid using Megazyme© Kit
- Preparation of reagent solutions (not supllied) :
- Add 200µL Buffer AL. Mix thoroughly by vortexing.
- Add 200µL ethanol (96%). Mix thoroughly by vortexing.
- Pipet the mixture into a DNeasy Mini spin column placed in a 2mL collection tube. Centrifuge at 6000g (8000rpm) for 1 minute. Discard the flow-through and collection tube.
- Place the spin column in a new 2mL collection tube. Add 500µL Buffer AW1. Centrifuge for 1 minute at 6000g. Discard the flow-through and collection tube.
- Place the spin column in a new 2mL collection tube. Add 500µL Buffer AW2. Centrifuge for 3 minutes at 20,000g. Discard the flow-through and collection tube.
- Transfer the spin column to a new 1.5mL or 2mL microcentrifuge tube.
- Elute the DNA by adding 200µL Buffer AE or sterile water to the center of the spin column membrane. Incubate for 1 minute at room temperature. Centrifuge for 1 minute at 6000g.
- Repeat this step for increased DNA yield.