Team:Paris Bettencourt/Protocols/Titration protocols for vitamin A with HPLC
Ferment It Yourself
iGEM Paris-Bettencourt 2O15
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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 protocols for vitamin A with HPLC- Suspend cells in 1 mL of sterile water.
- Add 0.5 to 0.75 mm glass beads and vortex for 3 minutes.
- Add 2.5 mL of 0.2% (wt/vol) pyrogallol dissolved in methanol and vortex for 3 minutes.
- Add 1.25 mL of 60% (wt/vol) KOH and vortex for 10 seconds.
- Incubate for 1 h at 75ºC, vortexing every 15 minutes.
- Add appropriate amount of hexane.
- Centrifuge tubes for 5 minutes at 2,800 rpm.
- Pipette 1 mL of hexane into a cuvette.