Now you are on the website of gene sequence, and the bottom paragraph is the sequence of exons 2 to 9. Generally, RT-PCR primers need to cross exon boundaries, which can improve the specificity.

You can copy the sequence of exon 2 (2 16...289) first.

atgga ggagccgcag tcagatccta gcgtcgagcc cctcttgagt caggaaacat tttcagacct atggaaact

Then copy the sequence of exon 3 (407...428).

actt cctgaaaaca acgttctg

Connect these two paragraphs.

atgga ggagccgcag tcagatcta gcgtcgagcc cctcttgagt caggaaacat tttcagacct atggaaactt cctgaaacaa acgttctg

Then go to the following web page to design primers.

http://www.ncbi.nlm.nih.gov/tools/primer-blast/

Paste the sequence you just spliced into the box at the top, and then click the button at the bottom. Click "Check" later, and you will get several sets of primers. At the top of the result is the location map. Just make sure that your reserve primers span the junction of sequence splicing.

I made this for you.

http://www . NCBI . NLM . NIH . gov/tools/primer-blast/primer tool . CGI？ ctg_time= 1397236878。 job _ key = Ahn QL 9 wld qfkle 4 glw 58 xjqjtk 8 NPF NK & amp； Check Status = Check