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

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.