If this gene does undergo alternative splicing, and the gene products do form proteins as you suggest in the future directions, what would that indicate?
If the products form proteins, it would be an confirmation that the gene is truly undergoing alternative splicing. We can then knock out that product and observe the effects on the cell, and find where these product are being used within the cell.
I made my decision through the use of the program Primer3Plus, which gave me numerous possible sets to use with different parameters. I picked three sets that had the best fit to run with my experiment, then input them into the program Benchling to see if they properly annealed to an exon. Two of them didn’t anneal correctly or at all to the gene, thus leaving me with my chosen primers.
I didn’t expect them to anneal at 201 bps, that was the size of the product that I predicted to be made with my primer set. The very light band of the gene specific primer well with no damaged DNA is right around that length.
If this gene does undergo alternative splicing, and the gene products do form proteins as you suggest in the future directions, what would that indicate?
If the products form proteins, it would be an confirmation that the gene is truly undergoing alternative splicing. We can then knock out that product and observe the effects on the cell, and find where these product are being used within the cell.
How did you decide what primers to initially use when you designed the gene specific primers?
I made my decision through the use of the program Primer3Plus, which gave me numerous possible sets to use with different parameters. I picked three sets that had the best fit to run with my experiment, then input them into the program Benchling to see if they properly annealed to an exon. Two of them didn’t anneal correctly or at all to the gene, thus leaving me with my chosen primers.
Why did you expect them to anneal at 201KB?
I didn’t expect them to anneal at 201 bps, that was the size of the product that I predicted to be made with my primer set. The very light band of the gene specific primer well with no damaged DNA is right around that length.