Hi everyone, I'm a lowly undergrad doing research underneath a Professor. Could someone help me out and tell me where I am going wrong? The professor told me she would always reply in a timely manner but this can mean anywhere from an hour to a day, and time is money friends!
I went to genome browser and looked specifically for the Mouse Gene TNFSF10, and copied the refseq into word. I then found the beginning primer at 1 to 263 nucleotides. I found the next exon, the reverse primer, at 8905-9043. So, I took the refseq into ncbi's primer blast tool. The settings of importance I think I should mention are: 2GC clamp (I require either 2 or 3 of these in primers) and primer must span exon-exon junction.
I'm also looking for primers with no more than 200 nucleotides in length, so unless if I misunderstood something which is very likely, how do I make it so my reverse primer isn't so big?
Thank you, and sorry for this mess.