Project:Sex typing with amelogenin/results-20120311
Two separate DNA extraction protocols: Chelex and meat tenderiser.
Chelex: rinse mouth with salt. Spit small amount of liquid into tube. Add 500ul Chelex. Incubate at 98 degrees for 6 minutes. Centrifuge for 5 minutes. Take 10ul supernatant.
Meat tenderiser: rinse mouth with salt. Spit small amount of liquid into tube. Add equal amount of detergent. Vortex for 30 seconds. Add a few grains meat tenderiser. Vortex for 30 seconds. Allow to sit for several seconds. Add a small amount of rubbing alcohol. Observe DNA strands precipitating out of solution. Take 10ul of liquid and DNA strands. Incubate at 70 degrees for several minutes. Add 5 ul water.
PCR was set at D=94 degrees, 1 minute; A=57 degrees, 1 minute; E = 72 degrees, 1 minute. I used 5 ul of sample, 6 ul of primers, 12.5 ul of Taq master mix.
Result was visualised on 1% agarose gel stained with 2ul/40ml of ethidium bromide under 300nm UV. DNA ladder was visible. Both samples showed a faint blur running ahead of the ladder and no other information.
I conclude that the DNA extraction technique is inadequate.