Zinc Finger Gene In Sea Lion

To observe a particular trait (zinc finger gene) in sea lion a tissue sample was obtained. To observe the presence on this gene the DNA was first extracted from the tissue sample. First a tissue sample was obtained in a 1.5 ml tube and the tube number, AK24667, was recorded. To this sample, 180 microliters of buffer ATL was added. After adding 20 microliters of proteinase K to the 1.5 ml tube, the sample was vortexed for 30 seconds to disrupt the pellet. The sample was then incubated at 56°C to lyse the tissue. The sample was checked every fifteen minutes and vortexed between each checking for an hour and a half until the tissue was completely lysed. The tissue sample was then again vortexed. Next 200 microliters of buffer AL was added and

In order to do this a PCR was set up. A PCR tube was obtained from the instructor. The PCR bead was then dissolved in 22 microliters primer cocktail. After the PCR bead was dissolved, three microliters of the DNA extracted from the first part of the experiment were added to the PCR mixture. This PCR mixture and the 1.5 ml extracted DNA samples were then submitted to the

To begin the process, a pre-prepared 1% agarose gel was obtained. The gel chamber was set up by placing the agarose gel into the chamber and submerging it in plentiful TAE buffer. The wells were filled with both PCR and DNA and shared between six students. The wells were labeled 1-7. The first well was pre-loaded with DNA ladder and labeled as 1 microliter kb DNA ladder. Next one microliter of DNA was mixed with one microliter of loading dye using a pipettor and loaded into the well. The same mixing process was completed for the PCR product, using one microliter of PRC product and one microliter of loading dye. For the purposes of this experiment, the DNA product was loaded into well six and the PCR product was loaded into well seven. Initially DNA was loaded into well five, however gel was pierced so samples were moved one well to the right. The gel was run at 100 V for one hour. The gel was stopped and observed under a UV illuminator. After the verification of both a single band in the DNA and PCR wells, the PCR product was to be