Alu PV92 Detection by PCR

Sip 10 mL of 0.9% saline solution and rinse your mouth vigorously for 30 seconds while gently chewing the inside of your cheek to release cells. Expel the saline back into the cup or tube.

Pipette 1 mL of mouth rinse into a 1.5 mL microcentrifuge tube, place in balanced configuration, and centrifuge at maximum speed for 2 minutes (or 5 minutes at 2,000 G). Locate the white cell pellet approximately the size of a match head; repeat if pellet is too small.

Pour off the saline solution carefully without losing the pellet, leaving a small amount at the bottom. Resuspend the pellet by rubbing the tube over a tube rack, vortexing, or flicking to break up all clumps.

Using a micropipette set to 20 microliters with aerosol barrier tips, transfer all resuspended cells to a screw cap tube containing InstaGene Matrix. Multiple transfers may be required; screw the cap on tightly and vortex the tubes.

Incubate the screw cap tubes at 56°C for 10 minutes, vortexing or shaking at the 5-minute mark. Remove and vortex the tubes, then incubate at 95°C for 5 minutes to inactivate the lysis buffer.

Vortex the tubes and centrifuge at top speed for 5 minutes (or 10 minutes at 2,000 G) to pellet debris. The DNA template is now ready for PCR or can be stored at 4°C for up to one month.

Pipette 20 microliters of PCR Master Mix into each labeled PCR tube, then add 20 microliters of DNA template from the corresponding labeled sample, avoiding the InstaGene Matrix pellet at the bottom. Pipette up and down to thoroughly mix.