Difference between revisions of "Team:Paris Bettencourt/Protocols/titration-acid-phytic"

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     <td><b>Titration of phytc acid using Megazyme© Kit</b><br />
 
     <td><b>Titration of phytc acid using Megazyme© Kit</b><br />
 
     <ul>
 
     <ul>
−
       <li>Preparation of reagent solutions (not supllied) :</li>
+
       <li><b>Preparation of reagent solutions (not supllied) :</b></li>
 
       <li>Add 200µL Buffer AL. Mix thoroughly by vortexing.</li>
 
       <li>Add 200µL Buffer AL. Mix thoroughly by vortexing.</li>
 
       <li>Add 200µL ethanol (96%). Mix thoroughly by vortexing.</li>
 
       <li>Add 200µL ethanol (96%). Mix thoroughly by vortexing.</li>

Revision as of 14:20, 28 August 2015

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.