Sydney Ortenberg

March 1, 2019

Lab 7: PCR Amplification

Objective/Purpose:

The objective of this lab was to modify our DNA sample in order to be used for PCR in next weeks lab. The purpose of this lab was to create our DNA for PCR so we can meta barcode our ciliate DNA to be used for the scientific community.

Procedures:

1. Wear gloves and clean the desk and work area with 10% bleach to sterilize the environment.
2. Place an empty tube, water, DNA sample, and V4 Primer in an ice bath.
3. Dilute DNA Sample if necessary  (in our case, we didn’t have to)
4. Take two tubes which have already containing 12.5ul 2X Taq Mix (one for control and one for treatment)
5. For the control tube, add 1ul primer and 11.5ul water and centrifuge for 15 seconds.
6. For the treatment tube, add 1ul DNA sample, 1ul primers, 10.5ul water and centrifuge for 15 seconds.
7. place in tube rack with other classes’ and label where you placed your tube on the sheet of paper.

Data:

Storage:

The ice bath and its contents of primer and water were taken back to the right area. The newly created DNA tubes were placed in the class storage rack and labeled where our group was placed. All other scientific equipment was disposed of and put back into the correct spot.

Conclusion and Future Steps:

Overall, this lab was successful and my group created this new DNA sample with ease. My group was lucky in that we did not have to dilute our DNA sample because our concentration was 96.3nm and that was close enough to 100ng. My group and I paid attention to small details so there would not be any mistakes or contamination of our DNA amplification tubes. I am looking forward to performing the PCR and what kind of results we get. It will be interesting to see what information we can take from our DNA sample and how far this whole experiment has come. I am thinking about what our poster will look like and how we will design it, and I am also starting to prepare for my midterm exam.