论文部分内容阅读
高分辨率熔解曲线分析(High resolution melting analysis,HRM)可以检测单碱基改变引起的DNA双链熔解温度(Tm)值变化,从而可以对样本在单核苷酸多态性分子标记(Single nucleotide polymor-phism,SNP)上进行基因分型。通过分析NCBI数据库中宽皮柑橘的表达序列标签(Expressed sequence tag,EST)数据鉴别SNP位点,并用小片段扩增法高分辨率熔解曲线分型技术(High resolution melting analysis of small amplicons)分析11个宽皮柑橘(Citrus reticulata)品种以及柳橙(Citrus sinensis Osbeck var.‘Liucheng’)的5个SNP位点的基因型。结果显示,小片段扩增法高分辨率熔解曲线分型可以快速、清楚地分辨纯合与杂合基因型,在校正温度差异后也可以很好地分辨同一个SNP位点不同的纯合型。统计分析表明样本在所有SNP位点上均存在多态性,5个SNP位点的平均多态性信息含量(PIC)为0.3190,显示样本在这组SNP位点上具有较高的杂合率。
High-resolution melting analysis (HRM) can detect single-base DNA double-strand melting temperature (Tm) changes in value, so that samples can be single nucleotide polymorphism in molecular markers (Single nucleotide polymor-phism, SNP). The SNP loci were identified by analyzing the expressed sequence tags (ESTs) of the mandarin orange in the NCBI database and analyzed by high resolution melting analysis of small amplicons Genotypes of five Citrus reticulata cultivars and five SNP loci of Citrus sinensis Osbeck var. ’Liucheng’. The results showed that the high-resolution melting curve typing of small fragment amplification method can quickly and clearly distinguish between homozygous and heterozygous genotypes, and can also well distinguish homozygous homozygotes of the same SNP site after correcting for temperature differences . Statistical analysis showed that the SNPs were polymorphic at all SNP loci. The average polymorphic information content (PIC) of the five SNPs was 0.3190, indicating that the samples had higher heterozygosity at the SNP loci .