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根据不同长度或不同碱基序列的DNA片段的熔解温度(Tm)不同的原理,将酶切扩增多态性序列(cleaved amplified polymorphic sequence,CAPS)与熔解曲线(melting curve,MC)技术相结合,建立了CAPS-MC技术,从而实现SNP分子标记基因分型。通过菜薹‘L58’和紫菜薹‘ZCT095’的基因组重测序数据与白菜基因组的参考序列(‘Chiifu-401-42’的基因组序列)的比对,预测了719个HindⅢ内切酶识别碱基序列的SNP位点,并将其设计成CAPS-MC分子标记,从中随机选择10对标记进行体系建立与优化,结果显示本体系有灵敏度强、精准性高、快速、高通量、经济实用等优势。
The cleaved amplified polymorphic sequence (CAPS) is combined with the melting curve (MC) technique according to the principle of different melting temperature (Tm) of DNA fragments of different length or different base sequences , Established CAPS-MC technology, in order to achieve SNP molecular marker genotyping. The alignment of 719 HindIII endonuclease recognition bases was predicted by comparing the genome reanimation data of the Chinese cabbage ’L58’ and the seaweed ’ZCT095’ with the reference sequence of the Chinese cabbage genome (’Chiifu-401-42’) Sequence of SNP sites, and designed it as CAPS-MC molecular marker, randomly selected from 10 pairs of markers to establish and optimize the system, the results show that the system has high sensitivity, high precision, fast, high throughput, economical and practical Advantage.