Biotech Lab Test Notes

University:
University of California, San Diego
Course:
AWP 3 | Analytical Writing
Academic year:

2024
Views:

247

Pages:

2
Author:

customer-8717572

1. Isolate the DNA from Strawberry a. Make lysis buffer (too break down cell membrane and cell wall) i. (90mL water, 1mL detergent, 0.5grams salt, then make up vol. to 100mL? Idk if its right) b. Mash up the strawberry with 10 mL of lysis buffer c. Filter d. Take 400microLiter of juice e. Put 2x of Isopropanol (800x) *precise amount is very important* f. Weigh mass g. Centrifuge (remember balance) for 10 minutes h. Dump the supernatant (liquids) i. Wait for it to dry ( j. Wash the pellet with 1mL of 70% Ethanol (to clean up the dna pellet better) k. centrifuge/ spin again l. Dump the supernatant (liquids) m. Resuspend pellet in 50microLiter of water n. Show Dr. Ghosh the pellet 2. DNA from cheek cells a. Lysis buffer b. Take 400microLiter of Lysis buffer c. Take toothpick and take cheek cells with blunt end 30s d. Swirl in lysis buffer (dispose toothpick)bhjm, e. Vortex for 1 minute (for animal cells most, no cell wall, further break down) f. 2x Isopropanol g. Weigh and centrifuge h. Dump the supernatant i. Wait for it to dry j. Wash the pellet with 1mL of 70% Ethanol k. centrifuge/spin again l. Dump supernatant and resuspend in 50microLiter of water m. Show Dr. Ghosh the pellet 3. Making/running a gel a. Make 1x TAE buffer (300mL for each gel) from the 50x TAE i. V(1)C(1) = V(2)C(2) b. Take 0.5gram of agarose in 50mL of 1x TAE c. Microwave 30sec, x2 d. Wait for cool (warm to touch) e. Put 5microLiter of gel stain f. Pour the gel, let it solidify for 15 minutes g. Take DNA (50microLiters), take 10 microLiters from that, add 2microLiter of gel loading dye h. Load the DNA + 1 Kb control DNA i. Run the gel at 80 V(volts) for 15 min j. Check under UV transilluminator k. Take a photo of gel