DNA Extraction for college laboratory setting
Label	one tube for	each	plant.
Harvest 2-3 seedlings and place in a mortar.
Fill with about 50 ml of liquid nitrogen.
Grind tissue with pestle.
Add 1 ml of extraction buffer to the tube.
Add 120 µl of 10% SDS.
Mix by inverting.
Incubate tube(s) at 65 ˚C for 20 minutes.
Add 300 µl 5M KOAc.
Mix well by inverting several times (important!
), then place on ice 5 minutes.
Centrifuge	for	5	minutes	at	>12,000	rpm.
Label	a	second	tube.
Pass 700 µl of the supernatant through a miracloth funnel into the second tube.
Add 600 µl of isopropanol.
Mix the contents thoroughly by inverting.
Spin	for	5	minutes	at	14,000	rpm.
Carefully pour off and discard the supernatant.
Use a P20 set to 20 µl to remove the remaining drops of liquid without disturbing the DNA pellet.
Add 500 µl of 70% ethanol and flick the tube until the pellet comes off the bottom.
Spin	5 minutes.
Pour off the ethanol.
Use a P20 set to 20 µl to remove the remaining drops without disturbing the pellet.
Leave	the	tube	open	on	the	bench	to	air dry	for	5-10	minutes.
Resuspend the DNA in 50 µl TE and incubate at room temperature for 5 minutes for complete resuspension.
Samples should be frozen for storage.
