From the design primer in each software, I choose primer design from three software. First pair http://www.bioinformatics.nl/cgi-bin/primer3plus/primer3plus.cgi 5’TTAGGCTTCATGTCCCTGCT 3’ and 5’CCCCTGGTTGTGAAATCAAT 3’, second software http://biotools.umassmed.edu/bioapps/primer3_www.cgi 5’TTAGGCTTCATGTCCCTGCT 3’ and 5’CCCCTGGTTGTGAAATCAAT 3’, and third http://www.ncbi.nlm.nih.gov/tools/primer-blast/ 5’GAAGTCGTGTGGCAACAACC 3’ and 5’ACGAGCTGTCCAACACTGAG 3’,Because there are have Primer Melting Temperature (Tm) value between 52-60 oC. GC content, The GC content of primer should be 40-60%.and GC content give indicate for Tm value. And there are low self complementary.