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解锁FFPE样本更多测序信息,他们都说好!
Pacbio SMART sequencing. “Genomic sequencing of DNA from FFPE using truXTRAC and Adaptive Focused Acoustics by Covaris, Inc. is compatible with single molecule sequencing.” Jeff Conroy, Chief Scientist Officer, OmniSeq “By using the Covaris truXTRAC FFPE DNA and RNA kits, OmniSeq testing requires less tissue and results infewer specimen failures when performing comprehensive genomics profiling on solid tumors.” Manu Gupta, University College London “Having compared various methods for DNA extraction from FFPE material, we now use Covaris followed by anautomated bead-based purification methodology. This has now become a standard practlce in our laboratories.” 如果你也苦恼于FFPE 核酸提取质量不高,那么Pacbio SMART单分子测序和Ominiseq都推荐的Covaris truXTRAC FFPE核酸提取解决方案你很有必要了解一下!点击下方小视频,简单了解下吧! Covaris FFPE 提取优势 可自动化 高得率 可DNA/RNA 共提 RNA 高DV200值 低样本起始量 低QNS率(无法诊断样本) 无需有机溶剂脱蜡 高测序覆盖度 超声脱蜡水解彻底 改进融合基因的检测 简单标准可自动化的提取流程! 样本收集到核酸纯化<3h,同一样本,可同时提取DNA和RNA,简单化操作流程,纯化可选柱纯化和自动磁珠富集,易于自动化规模化;可选NGS片段化步骤,核酸提取到片段化全程单管操作,无需转移,减少污染与样本损失。 AFA 超声脱蜡更彻底 还在用二甲苯、矿物油等脱蜡?其实二甲苯不仅对生物党们身体有毒,而且脱蜡又不彻底,稍微厚一点的(>10μm)组织切片就难以渗透,脱蜡效率就更低了。相信很多人要重复脱两遍蜡的时候。蜡脱不彻底会咋样?石蜡就成了阻碍阻碍下一步蛋白酶K消化的屏障,所以组织消化也不够,此外,二甲苯还容易造成核酸降解,本就量少的FFPE样本其实这么一折腾更后的核酸得率质量都受很大影响,也许很多意想不到的基因信息就这么不经意的丢失了。 Covaris技术 AFA超声来对FFPE样本进行超声乳化脱蜡,想象一下,有聚焦超声的帮助,裂解液和蛋白酶K能够由内而外、由外向内的全方位无死角侵入组织内部脱蜡乳化、组织消化,这样纳米级别的处理当然使得样品脱蜡消化更彻底! AFA超声脱蜡前后对比 更高的RNA DV200值 illumina 推荐用大于200nt RNA片段所占比值即DV200值来评估用于illumina文库构建的RNA样本质量,DV200值小于30%的样本不推荐用于文库构建,随着DV200值越高,RNA input量可以减少。从下图可以看出,使用Covaris 提取的FFPE样本RNA DV200值均符合建库要求。 Figure 1. Illumina uses 30% DV200 cut-off. DV200: Distribution Value 200, which is the percentage of RNA fragments larger than 200 nucleotides. (Levy, S. The ORIEN Project- Large Scale Integration of FFPE Material into a Multisite Research Alliance. Oral presentation at AACR 2019; April 1, 2019) 更高的测序覆盖度 下图为FFPE肾脏样本与新鲜样本提取DNA全基因组测序覆盖度比较:其中横坐标表示染色体,浅色为覆盖度小于10x的区域,深色为覆盖度大于10x的区域,Covaris truXTRAC提取的样本,测序覆盖度更加均一,更接近新鲜的冰冻样本! Figure 2. Whole Genome Sequencing Results (Chromosome 19 Coverage) Comparison Covaris truXTRAC, Competitor and fresh frozen tissue.Sequencing track from Q FFPE kit (red), Covaris (green) extracted FFPE kidney DNA, and fresh frozen (indigo) samples were loaded on the IGV viewer, and the coverageanalyzed for chromosome 19. Coverage of >10x are indicated in dark colors, coverage of <10X are indicated in light colors. Chromosomal view of coverage indicates that Covaris extracted DNA quality resembles that of the DNA extracted from fresh frozen tissue. 想要了解更多Covaris truXTRAC FFPE 信息可点击以下链接: 基因有限公司作为Covaris公司中国区独家代理商,为您提供更完整的DNA 片段化解决方案,也可为您提供ChIP实验,FFPE样本核酸提取解决方案等。基因有限公司的宗旨是“专才为专家服务”,有专业团队为各大科研机构提供安装培训,技术支持,应用培训与售后维护一条龙服务,赢得客户的一致好评与信任。想要了解更多,请关注我们的官方微信“基因快讯”或联系您身边的基因有限公司员工。