Rationale: The purpose of this lab is to complete DNA extraction and quantify the DNA with the Nanodrop.

Description of Procedures:

1. The titer of the lysate was determined to be 2.0 x 10^10.
2. 0.5 ml of sterile water was added to the DNA pellet to re-suspend it.
3. 2 ml of 37 C Clean Up Resin was added to the pellet and swirled.
4. Resin pellet was transferred to 2 microcentrifuge tubes and spun for 3 minutes at 12,500 xg.
5. Supernatant pulled off with a bulb pipette and 1 ml of 80% isopropanol was added to each tube. The tube was spun at 12,500 xg for 3 minutes.
6. Step 4 repeated
7. Supernatant pulled off again and 1 ml of 80% isopropanol added to each tube. Liquid transferred to a two column syringe and allowed the vacuum to pull the liquid through the column to separate the DNA.
8. Column added to a clean microcentrifuge tube and spun at 12,000 xg for 5 minutes.
9. Transferred to a clean tube and 80 ul of 80 C elution buffer added to the tube. It was then spun in the centrifuge at 12,000 xg for 1 minute.
10. DNA was quantified using the Nanodrop.

Observations/Results:

• DNA- 808.978 ng/ul
• A260/A280- 2.049
• A260/A230- 0.193

Interpretations:
The procedure was completed. The DNA was extracted and quantified. The A260/A230 value was low, meaning there could be too much guanidinium thiocyanate left in the DNA. The next step would be to perform PCR and run a gel, but there is not enough time left in the lab.

Extracted DNA and Spot Titer Test: