Lab 3 Notes

Directions for Lab 3: Plant gene sequencing 1. DNA extraction
1.) Select 2 plants/group
2.) Gather about 100mg (0.1g) of plant leaves
3.) Dice plant matter with scalpel blade
4.) Put diced plant in microcentrifuge tube and LABEL!
5.) Smush plant matter in microfuge tube with 200mL of Lysis Buffer, using a pestle
6.) Shake and add more Lysis Buffer and grind up some more until no visible particles
7.) Centrifuge at 16,800rpm for 5min.
8.) Transfer 400mL of supernatant into a new tube; add 500mL of 7% Ethanol. Then pipet up and down to mix.
9.) Transfer plant goo+E+OH(from #8) into the purple column(about 800mL)
10.) Centrifuge for 1 min. DON'T SPILL and DON'T CAP
11.) Discard flow thru liquid (keep tube)
12.) Add 800ml wash buffer to each column(purple thing) and centrifuge for 1 min. Discard the flow-thru.
13.) Repeat step 12 two more times.
14.) Centrifuge for 2min. to dry.
15.) Add 80mL of sterile H2O to the column and let it sit for 2min.
16.) Centrifuge column with new capped tube. KEEP FLOW-THRU(DNA sample).