CAPS与熔解曲线技术相结合的菜薹SNP基因分型体系的建立与验证

 根据不同长度或不同碱基序列的DNA片段的熔解温度（T_m）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。

Establishment and Validation of SNP Genotyping System by CAPS Combining Melting Curve in Brassica campestris

Melting temperature（T_m）was different according to the different lengths or nucleotide sequences of DNA fragments. In this report，we established a novel single nucleotide polymorphism（SNP）genotyping system refered as CAPS-MC approach. The CAPS-MC approach combined cleaved amplified polymorphic sequence（CAPS）with melting curve（MC）analysis. By aligning whole genome re-sequencing data from two Brassica campestris accessions，‘L58’and‘ZCT095’，to the reference genome sequence of‘Chiifu-401-42’，a total of 719 HindⅢ endonuclease recognition nucleotide sequence SNP polymorphisms were converted to CAPS-MC markers. We established and optimized CAPS-MC system by randomly selecting 10 pairs of these markers. Our results clearly demonstrated that CAPS-MC approach was sensitive，accurate，rapid and inexpensive. Because the melting curve of 96 samples can be measured in 22 mins once，it is well suited for high-throughput genotyping.