ALU PCR lab
Purpose:
To isolate our own DNA (from cheek cells) and create a PCR reaction to amplify the Alu insert in our genetic code
Materials:
0.9% saline solution
PCR tubes
1x TAE stock
Master Mix
Primer Mix
PCR tubes
Micropipette + tips
microcentrifuge + tubes
agarose
Gel chambers + molds
chelex solution
Water
Waste container
Tube racks
Procedure:
1) Wash mouth in 0.9% saline solution for 30sec. then spit solution (plus everything else) into cup
2) Pipette 1-1.5ml. into microcentrifuge tube. Label tube and spin for 1min.
3) Pour out supernatant and resuspend cell bead at bottom of tube
4) Pipette 50 uL into new tube with Chelex beads
5) Heat for 10min.
6) Spin for 1min, add 50uL of supernatant to new tube
7) New tube- add 20uL of Master and Primer mix + 10uL of DNA. Put in thermal cycler
8) Spin tubes, then add 20uL of DNA and 4uL of loading dye
9) Spin and load into gels
For Gels:
1) Mix 50 mL TAE with 1g agarose and heat until agarose dissolves
2) Wait for solution to cool and pour into mold. Remove comb when hardened.
To isolate our own DNA (from cheek cells) and create a PCR reaction to amplify the Alu insert in our genetic code
Materials:
0.9% saline solution
PCR tubes
1x TAE stock
Master Mix
Primer Mix
PCR tubes
Micropipette + tips
microcentrifuge + tubes
agarose
Gel chambers + molds
chelex solution
Water
Waste container
Tube racks
Procedure:
1) Wash mouth in 0.9% saline solution for 30sec. then spit solution (plus everything else) into cup
2) Pipette 1-1.5ml. into microcentrifuge tube. Label tube and spin for 1min.
3) Pour out supernatant and resuspend cell bead at bottom of tube
4) Pipette 50 uL into new tube with Chelex beads
5) Heat for 10min.
6) Spin for 1min, add 50uL of supernatant to new tube
7) New tube- add 20uL of Master and Primer mix + 10uL of DNA. Put in thermal cycler
8) Spin tubes, then add 20uL of DNA and 4uL of loading dye
9) Spin and load into gels
For Gels:
1) Mix 50 mL TAE with 1g agarose and heat until agarose dissolves
2) Wait for solution to cool and pour into mold. Remove comb when hardened.