摘要
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。
Melting temperature （Tin） 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 HindlII 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.
出处
《园艺学报》
CAS
CSCD
北大核心
2013年第10期1916-1926,共11页
Acta Horticulturae Sinica
基金
国家重点基础研究发展计划(‘973’计划)项目(2012CB113906)
