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