Week 10

This week brought the news that my research project was accepted for presentation at the Estrella Mountain Student Conference; the S-STEM scholars, including myself, will also be presenting at Metrotech high school on April 25th. I look forward to the opportunity to hone my research for presentation to both groups. Truth be told, I am more interested in presenting to the vocational students than the other undergrads, as I look at it as an opportunity to potentially open a student's mind to the possibility of higher education.

My initial work in the lab this week was focused on completing an extraction protocol on the E. coli DNA that I have been using during this first phase of my project. Then I used a Promega Wizard DNA clean-up kit to remove contaminants from all four samples of DNA that I extracted last week. The cleanup protocol is as follows:

1. Re-suspend clean-up resin by inversion.
2. Add 1 mL of the re-suspended resin to a 1.5 mL micro centrifuge tube.
3. Add the sample to the clean-up resin and mix by inverting several times.
4. Attach a micro column to the barrel of a sterile 5 mL syringe.
5. Pipette the clean-up resin containing the bound DNA into the syringe barrel.
6. Slowly and gently push the slurry into the mini column with the syringe plunger.
7. Add 2 mL of 80% isopropanol to the syringe and gently push the solution through the mini column.
8. Transfer the mini column to an eppendorf tube and centrifuge for 2 minutes at 13200X g.
9. Transfer the mini column to a new eppendorf tube and apply 50 µl of 1x TE buffer and wait one minute.
10. Centrifuge the mini-column for thirty seconds at 13200x g.
11. Discard the mini column and keep the eppendorf tube containing the supernatant. The prepared sample can be stored at -20°.

I then prepared all four samples for an electrophoresis to identify if DNA was extracted. I used a 1x TAE solution to prepare both the buffer and the gel, then added 2 mL of each sample separate wells, followed by 1 mL each of loading dye and SYBR green. After adding the buffer, I initiated the ep process and covered it with a lab coat to reduce light interference. The gel ran from 24 minutes. The following is an image of the completed gel, with the two wells to the left consisting of control DNA provided by the lab coordinator, Josh.