Purpose:
To use the DNA located on our cheek cells, and create a PCR reaction to amplify the ALU Insert
MAterials:
0.9% saline solution -micropipetts, tips
-waste container -microcentrifuge, tubes
-PCR tubes -agarose
-1x TAE -gel chambers + molds
-load dye -chelex
-racks -primer mix
-master mix -H2O
- (+,-) control DNA
-waste container -microcentrifuge, tubes
-PCR tubes -agarose
-1x TAE -gel chambers + molds
-load dye -chelex
-racks -primer mix
-master mix -H2O
- (+,-) control DNA
Procedure:
1. Swish the 0.9% saline solution, that is provided, for 30 secs then spit it out into a cup.
2. Move 1 mL - 1.5 mL of used saline solution into a microfuge tube, the spin it for 1 min.
3. Remove the supernatant from the tube, spin the cells for a couple of seconds to re-suspend them, move 50 uL of cells to a tube w/ chelex beads, then place the tube in the heat block for 10 min.
4. Use the centrifuge to spin the tube for 1 min. Move 50 uL of supernatant into a new microfuge tube, place in a rack.
5. Get a new tube, add 20 uL of the master mix, 20 uL of the primer mix , and 10 uL of your own DNA, then place the tube in the thermal cycler.
6. Give the tubes a quick spin in the centrifuge, Move 20 uL of the DNA sample to a new microfuge tube, add 4 uL loading dye, the spin the tube again.
7. Load 20 uL of the DNA sample into a gel well.
2. Move 1 mL - 1.5 mL of used saline solution into a microfuge tube, the spin it for 1 min.
3. Remove the supernatant from the tube, spin the cells for a couple of seconds to re-suspend them, move 50 uL of cells to a tube w/ chelex beads, then place the tube in the heat block for 10 min.
4. Use the centrifuge to spin the tube for 1 min. Move 50 uL of supernatant into a new microfuge tube, place in a rack.
5. Get a new tube, add 20 uL of the master mix, 20 uL of the primer mix , and 10 uL of your own DNA, then place the tube in the thermal cycler.
6. Give the tubes a quick spin in the centrifuge, Move 20 uL of the DNA sample to a new microfuge tube, add 4 uL loading dye, the spin the tube again.
7. Load 20 uL of the DNA sample into a gel well.
Gel procedure:
1. Mix 50 mL of 1x TAE w/ 1 g of agarose, heat until dissolved
2. Pore into mold when cooled
2. Pore into mold when cooled