I used primer1 tool for design primer for TCF7L2 SNP, rs12255372
but the result was not stastify.
http://primer1.soton.ac.uk/runprimer1.cgi?TGTCTGTTTCTGAGGAATCCTGGGATGGGGCCACTGGCTCTTTAAGAGAGAGCCCGATTTGGAAATCTAGGACTTGATTGTTGATTATGGGCAATAGATACATTTTAAGAATGATGTTGTAGGCTGTATGAAGTCATTTGATGATTGTTTTGTTAATGGCTTGCAGGTCAGATTTTCATCTTTTTAAATTAATTATCATAGAAGGAGAAAACAACTGGATTTCAGAATTGTCCCTTGAGGTGTACTGGAAACTAAGGCGTGAGGGACTCATAGGGGTCTGGCTTGGAAAGTGTATTGCTATGTCCAGTTTACACATAAGGATGTGCAAATCCAGCAGGTTAGCTGAGCTGCCCAGGAATATCCAGGCAAGAATGACCATATTCTGATAATTACTCAGGCCTCTGCCTCATCTCCGCTGCCCCCCCGCCCCCTGACTCTCTTCTGAGTGCCAGATTCAGCCTCCATTTGAATGCCAAATAGACAGGAAATTAGCATGCCCAGAATCCACGTCTTTAGTGCACTCTCTCCCCAGCTCCAAACCTGTTACTGCTTGTGTTCAACATCTCAGTAAAGCTCAACAACATCGACCCATTACTTAGGCCTCAAACCTTGGGTGGCATCGTCGATTGCTCTTTTCTTTCATACCCCACATTCAACCCATCAGCCCATCCCACAGGCCCAAGTGTGTCCTCTCTACCTTCAAAGCGTGTGTGGCATCCA+374+G+T+28+30+26+200+300+100+1.5+1.1+65+80+50+80+20+8.00+3.00+50+50+10
could you help me design this primer? and teach me some step.
but the result was not stastify.
http://primer1.soton.ac.uk/runprimer1.cgi?TGTCTGTTTCTGAGGAATCCTGGGATGGGGCCACTGGCTCTTTAAGAGAGAGCCCGATTTGGAAATCTAGGACTTGATTGTTGATTATGGGCAATAGATACATTTTAAGAATGATGTTGTAGGCTGTATGAAGTCATTTGATGATTGTTTTGTTAATGGCTTGCAGGTCAGATTTTCATCTTTTTAAATTAATTATCATAGAAGGAGAAAACAACTGGATTTCAGAATTGTCCCTTGAGGTGTACTGGAAACTAAGGCGTGAGGGACTCATAGGGGTCTGGCTTGGAAAGTGTATTGCTATGTCCAGTTTACACATAAGGATGTGCAAATCCAGCAGGTTAGCTGAGCTGCCCAGGAATATCCAGGCAAGAATGACCATATTCTGATAATTACTCAGGCCTCTGCCTCATCTCCGCTGCCCCCCCGCCCCCTGACTCTCTTCTGAGTGCCAGATTCAGCCTCCATTTGAATGCCAAATAGACAGGAAATTAGCATGCCCAGAATCCACGTCTTTAGTGCACTCTCTCCCCAGCTCCAAACCTGTTACTGCTTGTGTTCAACATCTCAGTAAAGCTCAACAACATCGACCCATTACTTAGGCCTCAAACCTTGGGTGGCATCGTCGATTGCTCTTTTCTTTCATACCCCACATTCAACCCATCAGCCCATCCCACAGGCCCAAGTGTGTCCTCTCTACCTTCAAAGCGTGTGTGGCATCCA+374+G+T+28+30+26+200+300+100+1.5+1.1+65+80+50+80+20+8.00+3.00+50+50+10
could you help me design this primer? and teach me some step.