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PCR Detection of Genetically Modified Foods: DNA Extraction, Part I (Begin…
PCR Detection of Genetically Modified Foods: DNA Extraction, Part I
Begin DNA Extraction of Dried Foods
If the food is a dried solid, crush it first. Wear gloves, transfer a few pieces into freezer bag. Seal bag and crush the food.
Weigh .1 g of each dried food and place in disposable microcentrifuge tube.
Save fresh fruits and vegetables for later in protocol
Add 200 uL of nuclease free water to the tube.
Obtain the 4 samples for testing
Homogenize each sample to slurry.
Slide lock onto each tube and incubate for 2 hours.
Add 860uL of extraction buffer, 100 uL of 5M Guanidine HCL and 40 uL of proteinase K
DNA Extraction of Wet Foods
Put into microcentrifuge tube.
Freeze the food by dropping the tube into the liquid nitrogen.
Cut small piece of food with razor
After 30 seconds, use scoop to remove the tubes from the liquid nitrogen.
Wash food's outside with tap water
Put the tubes in the rack in freezer for next time.
Don't start until Liquid Nitrogen is here
Continue DNA Extraction of Dried/Processed Foods
Allow to cool for 10 minutes.
Centrifuge 10 minutes at 14,000 x
Remove samples from waterbath
Pipet the top layer into a clean labelled microcentrifuge tube. Freeze until next period.
Shivani Bisen
TA: Daniel