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

Line 8: Line 8:
 
     <ul>
 
     <ul>
 
       <li><b>Preparation of reagent solutions (not supllied) :</b></li>
 
       <li><b>Preparation of reagent solutions (not supllied) :</b></li>
−
      <li>Add 200µL Buffer AL. Mix thoroughly by vortexing.</li>
+
          <ul>
−
      <li>Add 200µL ethanol (96%). Mix thoroughly by vortexing.</li>
+
          <li><i>Trichloroacetic acid  (50% w/v) : 100mL</i></li>
−
      <li>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.</li>
+
          <li>50g of thrichloroacetic to 60mL of distilled water and dissolve. Make to volume (100mL) with distilled water. (Stable for > 6 moths at 4°C)</li>
−
      <li>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.</li>
+
          <li></li>
−
      <li>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.</li>
+
          <li></li>
−
      <li>Transfer the spin column to a new 1.5mL or 2mL microcentrifuge tube.</li>
+
          <li></li>
−
      <li>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.</li>
+
          </ul>
−
       <li>Repeat this step for increased DNA yield.</li>
+
       <li>
 
     </ul>
 
     </ul>
 
     </td>
 
     </td>

Revision as of 14:29, 28 August 2015

Titration of phytc acid using Megazyme© Kit
  • Preparation of reagent solutions (not supllied) :
    • Trichloroacetic acid (50% w/v) : 100mL
    • 50g of thrichloroacetic to 60mL of distilled water and dissolve. Make to volume (100mL) with distilled water. (Stable for > 6 moths at 4°C)