reference:
https://www.sogou.com/link?url=hedJjaC291NYNxVe4xgB4c3bUxXRMqZrT93cntTAgYfyBbRAdP9kIA..
https://liucheng.name/457/
Download the official website:
https://webblast.ipk-gatersleben.de/misa/index.php?action=3&help=2 ## download misa
https://webblast.ipk-gatersleben.de/misa/index.php?action=3&help=3 ## download p3_in.pl p3_out.pl
1, the use of Misa: perl Misa .pl prefix.10k.fa ## FA sequence must have two points separated by generation, and have the same directory misa.pl parameter file misa.ini. Generate results prefix.10k.fa.misa , prefix.10k.fa.statistics
prefix.10k.fa.misa: List type and microsatellite loci in tabular form;
prefix.10k.fa.statistics: Type and number of microsatellites statistical frequency;
2, Perl p3_in.pl prefix.10k.fa.misa ## is adapted to increase the parameter prefix.10k.fa.misa primer3_core input format, and output is prefix.10k.fa.ssr.p3in
. 3, primer3_core run generated, home / renpp / biosoft / primer3 / primer3_core <prefix.10k.fa.ssr.p3in> prefix.10k.fa.ssr.p3out ## output is prefix.10k.fa.ssr.p3out
. 4, p3_out.pl of primers designed for processing, generates a natural read format perl p3_out.pl prefix.10k.fa.ssr.p3out prefix.10k.fa.misa ## generate a result prefix.10k.fa.ssr .p3out.primer.xls