Difference between revisions of "Team:NYU Shanghai/Protocols"

Line 321: Line 321:
 
  <br>
 
  <br>
 
  <div id="mpText" style="display:none">
 
  <div id="mpText" style="display:none">
−
    This is how you miniprep.
+
      Estimated time
 +
      <br><br>Materials
 +
      <li>Biomiga Miniprep Kit</li>
 +
      <br><br>Procedure
 +
      <ol>
 +
      <li>Harvest the bacterial culture by centrifugation for 1 min at 10,000 rpm. Pour off the supernatant and blot the inverted tube on a paper towel to remove residue medium. Remove the residue medium completely. </li>
 +
      <li>Add 250 µL Buffer A1 (Add RNase A to Buffer A1 before use) and completely resuspend bacterial pellet by vortexing or pipetting </li>
 +
      <li>Add 250 µL Buffer B1, mix gently by inverting the tube 10 times (do not vortex), and incubate at room temperature for 5 minutes. </li>
 +
      <li>Add 350 µL Buffer N1, mix completely by inverting/shaking the vial for 5 times and sharp hand shaking for 2 times. <br>Note: Incubating the lysate in ice for 1 min will improve the yield. </li>
 +
      <li>Centrifuge the lysate at 13,000 rpm for 10 minutes at room temperature. If the lysate doesn’t appear clean, reverse the tube angle, centrifuge for 5 more minutes and then transfer the clear lysate to DNA column. </li>
 +
      <li>Carefully transfer the clear lysate into a DNA column with a collection tube, avoid the precipitations, spin at 13,000 rpm for 1 minute, discard the flow-through and put the column back to the collection tube. </li>
 +
      <li>Add 500 µL Buffer KB into the spin column, centrifuge at 13,000 rpm for 1 minute. Remove the spin column from the tube and discard the flow-through. Put the column back to the collection tube. </li>
 +
      <li>Add 650 µL DNA Wash Buffer (Add ethanol to DNA wash buffer before use) into the spin column, centrifuge at 13,000 rpm for 1 minute at room temperature. Remove the spin column from the tube and discard the flowthrough.</li>
 +
      <li>Reinsert the spin column, with the lid open, into the collection tube and centrifuge for 2 minutes at 13,000 rpm. </li>
 +
      <li>Carefully transfer the spin column into a sterile 1.5 mL tube and add 50-100 µL (> 50 µL) Sterile ddH20 or Elution Buffer into the center of the column and let it stand for 2 minutes. Elute the DNA by centrifugation at 13,000 rpm for 1 minute. Reload the eluate into the column and elute again.</li>
 +
      </ol>
 
</div>
 
</div>
 
</div>
 
</div>

Revision as of 01:56, 15 September 2015

Protocols

Recipes

Making Color

3A Assembly