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Assignment -
The upper-case, blue letters are the 14th exon (of 20) in the Hephl1 gene in mice. The lower-case (black) letters are from the flanking introns. The highlighted bases indicate primers that may be used to generate an amplimer for this segment, using either wild (273 bp) or mutant (274 bp) genomic DNA as a template in a standard PCR. In the mutant mouse, an extra A is inserted at the position of the red star.
For our colony maintenance, we plan to continuously backcross cw carriers to B10 +/+, so all of the kids would be phenotypically normal (a mix of cw/+ and +/+ genotypes). We need determine which are the carriers before we can produce the next generation. Sequencing the amplimer by primer extension would work, but that would take most of a week to get done (since we would have to hire a contractor). Instead, can you design an endonuclease-sensitivity test that would allow us to get the answer ourselves?
Attachment:- Assignemnt File.rar
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The solution file has NEB cutter results and attached files have list of flanking ends sensitive compatible to the ends., Restriction endonuclease test was performed and results attached
thank you very much for your response. I do appreciate it. I will make the payment right now. can I have the solution with explanation since I am a very new graduate student? please is there any chance that the work will be done by today 4:30 pm ?? it will be awesome. this assignment is about finding as much as we can of restriction enzymes(cutter). we can use the website he mentioned in the assignment. Fortunately the solution is correct. It is to find the restriction enzymes that cut in one genome and doesn’t cut in the mutant genome. Or vice versa. To be able to differentiate between both of them.The assignment is to find the cutter that will cut the mutant gene after adding A in the star site and nit cutting the wild one and then run it in the gel electrophoresis. Thanks. Nicely prepared work. It is clear and amazing work as per the requirements, all the points are covered. I would definitely be coming back with new assignments. Thank you so much for all the efforts you and your team has put into completing this assignment for me.
This assignment is due on . Please submit your experimental recommendations in a word doc, typed double-spaced. Sequencing the amplimer by primer extension would work, but that would take most of a week to get done (since we would have to hire a contractor).
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