Team:UNC Chapel Hill/1 July 2009

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'Frozen Stock' and 'Mini-Prep' in Scott's Lab (6/30 - 7/1) Part II

Notes

  • Peter and Pavel created a frozen stock of the bacteria transformed with [http://partsregistry.org/Part:BBa_I13521 RFP].

Procedure to create a frozen stock

  1. Add 750µL of bacteria and 250µL of 60% glycerol into a centrifuge tube.
  2. Shake it well to mix.
  3. Label the tube with colored tape.
  4. Put in into the -80 degree C freezer iGem box (2nd row from top, first drawer from left)

Mini-Prep (Protocol pamphlet is available in the lab as well)

  1. Add 1.5µL of bacteria into a small 1.5µL centrifuge tube and label.
  2. Spin in the centrifuge on the highest speed (14,000 rpm) for 1 min.
  3. Add 400µL of lysis.
  4. Put on vortexer until homogenous (~30 sec)
  5. Let sit for 3 min
  6. Spin again for 1 min at 14,000 rpm.
  7. Add 400µL of wash buffer
  8. Spin again for 1 min
  9. Take bottom off and pour out the liquid.
  10. Centrifuge again
  11. Warm up a bottle of elution buffer in microwave for 10 sec
  12. Add 50µL of elution buffer to the column
  13. Spin again
  14. What is left is the DNA. Put into box in -20 degree C freezer.
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