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首页> 外文期刊>BMC research notes >Novel approach for deriving genome wide SNP analysis data from archived blood spots
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Novel approach for deriving genome wide SNP analysis data from archived blood spots

机译:从存档的血斑中提取全基因组SNP分析数据的新方法

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Background The ability to transport and store DNA at room temperature in low volumes has the advantage of optimising cost, time and storage space. Blood spots on adapted filter papers are popular for this, with FTA (Flinders Technology Associates) Whatman?TM technology being one of the most recent. Plant material, plasmids, viral particles, bacteria and animal blood have been stored and transported successfully using this technology, however the method of porcine DNA extraction from FTA Whatman?TM cards is a relatively new approach, allowing nucleic acids to be ready for downstream applications such as PCR, whole genome amplification, sequencing and subsequent application to single nucleotide polymorphism microarrays has hitherto been under-explored. Findings DNA was extracted from FTA Whatman?TM cards (following adaptations of the manufacturer’s instructions), whole genome amplified and subsequently analysed to validate the integrity of the DNA for downstream SNP analysis. DNA was successfully extracted from 288/288 samples and amplified by WGA. Allele dropout post WGA, was observed in less than 2% of samples and there was no clear evidence of amplification bias nor contamination. Acceptable call rates on porcine SNP chips were also achieved using DNA extracted and amplified in this way. Conclusions DNA extracted from FTA Whatman cards is of a high enough quality and quantity following whole genomic amplification to perform meaningful SNP chip studies.
机译:背景技术在室温下以小体积运输和存储DNA的能力具有优化成本,时间和存储空间的优势。适应滤纸上的血斑很受欢迎,其中FTA(Flinders Technology Associates)Whatman? TM 技术是最新的技术之一。使用该技术已成功地存储和运输了植物材料,质粒,病毒颗粒,细菌和动物血液,但是从FTA Whatman提取猪DNA的方法是 TM 卡是一种相对较新的方法,迄今为止,核酸尚未准备好用于下游应用,如PCR,全基因组扩增,测序以及随后应用于单核苷酸多态性微阵列。从FTA Whatman? TM 卡中提取发现的DNA(按照制造商的说明进行改编),扩增全基因组,随后进行分析以验证下游DNA的完整性SNP分析。从288/288样品中成功提取DNA,并通过WGA扩增。 WGA后,不到2%的样品中发现等位基因缺失,没有明显的证据表明扩增偏差或污染。使用以此方式提取和扩增的DNA,也可以达到猪SNP芯片可接受的调用率。结论从FTA Whatman卡提取的DNA在整个基因组扩增后具有足够高的质量和数量,可以进行有意义的SNP芯片研究。

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