Mitochondrial DNA

Lab 7- Mitochondrial DNA

Note: for this activity you just will need to complete the chart shown at the end of this document and type or paste your answers into the space provided on moodle. *Note: The mt control region was selected for this lab because it is phenotypically neutral- it does not encode protein and has no known relationship to disease states, sex determination or any other phenotype. Nonetheless, for your own privacy we will label all samples only by a number that you’ll get as you begin the extraction. Only those numbers will be sent to the company that is sequencing this short region of your DNA, not your names. Sequence Analysis In this lab, you would have amplified your own DNA, and you will now compare it to known sequences on the DNA learning center server. *Note: The mt control region was selected for this lab because it is phenotypically neutral- it does not encode protein and has no known relationship to disease states, sex determination or any other phenotype. Nonetheless, for privacy reasons I labeled all samples only by their assigned number, and that is all that Genewiz has connected to your DNA samples. Protocol: 1. Access the DNAsubway website as directed in class. http://dnasubway.iplantcollaborative.org/ Either “Enter as Guest” or if you want you can Register to set up a free account. The benefit of the account is if you work on your sequences at one point you can come back to it later, otherwise you will have to start all over if you want to stop and continue your work later. 2. Click on the blue line, “Determine Sequence Relationships.” Click on Phylogenetics: mtDNA, then choose Import trace files from DNALC. Type in my last name in the search box and choose the most recent option. Click on the tracking number, and you should see the sequences- they should be named something like “KO1-LCMvF_A01.” KO because Sister Susan also sequenced some samples for her class, and LCMvF for the primer used for sequencing. Choose select all and “Add Selected Files.” Name your project and give it a description. 3. You’ll now see a blue subway line along the top of the page, and at each “Stop” you’ll see a blue circle, and then a line down to various smaller circles. This shows you the steps you need to complete. 4. Under “Assemble Sequences” click on “Sequence Viewer.” This will show you all the sequences. Notice that after you do this, the R turns to a V, meaning you have viewed/completed this step. 5. Click on “Sequence Trimmer,” which will remove the ends where the sequences was not sure about the sequence. 6. Ignore “Pair Builder” and “Consensus Builder” at this point, we only did the forward sequence here but if you did a reverse sequence of your same sample you would use this feature to match the two results.

Don't use plagiarized sources. Get Your Custom Essay on
Mitochondrial DNA
Just from $13/Page
Order Essay