SSR marker development for curcuma kwangsiensis

The searching principle was that only di-, tri-, tetra-, penta- and hexa-nucleotides with a minimum of 8, 5, 5,4 and 3 repeats will be considered as SSR loci. The primers of each EST-SSRs were designed using Primer Premier 5.0 (Premier Biosoft International, CA, USA) with following criterias: PCR fragment size- 100-350 bp; primer length- 16-24 bp; Tm- 55-65 °C; GC content- 40-60 %.

本研究的筛选原则为：仅当二核苷酸、三核苷酸、四核苷酸、五核苷酸与六核苷酸的重复次数分别不低于8次、5次、5次、4次与3次时，方可被认定为简单重复序列（Simple Sequence Repeats，SSR）位点。随后，利用Primer Premier 5.0软件（美国加利福尼亚州Premier Biosoft International公司）对各表达序列标签-简单重复序列（Expressed Sequence Tags - Simple Sequence Repeats，EST-SSR）设计引物，设计标准如下：PCR扩增产物片段长度为100~350 bp；引物长度为16~24 bp；解链温度（Tm）为55~65 ℃；GC含量为40%~60%。