Read through the assignment carefully before you start!! It will make answering the questions easier!!Īdd your name and e-mail address on top of the page!!!Ī family with the STXBP1 mutation requested that you make a model mouse in order to test various treatment options, and to study the nature of the disease. Your use CRISPR to introduce the mutation, and due to the distance, have to add several silent muations as well. The first thing you do is sequence the region in mouse to make sure the wild type is the same as the published sequence. You set up a PCR and send it to the sequencing unit with the forward primer to ensure that your sequence is correct. Ideally, we would send it with both forward and reverse primers, but due to primer design considerations, in this case, we wouldn't be able to see the change in the reverse primer. You receive an email that your sequence is waiting - now you have to download. The sequences are in the moodle assignment folder (if you can't get in, send me an email)Ĭlick the link on the main moodle course page: Sequences for Assignment 2. Open the DNASeq folder, there should be two subfolders inside: On a PC, you have to extract the files (it downloads as a zipped file) On a Mac, it will either expand automatically, or you double click to expand. Go into the wildtype sequence folder, and double click on the file ending in. Now we are going to look at the chromatograms.ĭownload a demo copy of Sequencher, unless your lab has a licensed copy, in which case you can use the "real" thing. If you are in Midrasha classroom B, it is already installed.Ĭlick on the link and there is a button for download. From the file menu, choose "Import - Sequences" and choose the. ab1 files from both of the sequence folders (one at a time). /rebates/&252fgeany-kosub-bradley. You now have a project with two sequences in it. #Dna sequencing sequencher heterozygous mutations download#.
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