RNA-seq可分为单端测序(Single-end sequencing)和双端测序(Paired-end sequencing)。对于geneA而言,单端测序表示仅沿着Read1方向(正向)开始测序,最后回帖到参考基因组上的只有1条reads,由此统计的值称为reads count。双端测序表示先沿着Read1方向测序,再从Read2方向开始测,最后回帖到参考基因组上有2个reads,这2条re...
第二代测序技术称为高通量测序(High-ThroughputSequencing),又名下一代测序(Next Generation Sequencing NGS)。顾名思义,它们解决了第一代测序中的低通量的缺陷,同时大大降低测序成本,目前使用最广的是illumina公司的Solexa,Hiseq技术,其核心技术大致相同,介绍如下-- 步骤一:构建文库 目的:在需要测序的DNA片段两端加...
基于Illumina高通量测序平台的RNA-Seq技术能够在单核苷酸水平对任意物种的整体转录活动进行检测,在分析转录本的结构和表达水平的同时,还能发现未知转录本和稀有转录本,精确地识别可变剪切位点以及cSNP(编码序列单核苷酸多态性),提供最全面的转录组信息。测序深度(Sequencing Depth)指测序得到的碱基总量与目标测序片段大小的比...
1. Moreover, the combination of RNA-seq and re-sequencing can be used both to remove false positives when inferring fusion genes and to analyze copy number alterations. 2. The statistically significant correlations that were observed, however, accounted for relatively small effects. (DNA methylatio...
1. Moreover, the combination of RNA-seq and re-sequencing can be used both to remove false positives when inferring fusion genes and to analyze copy number alterations. 2. The statistically significant correlations that were observed, however, accounted for relatively small effects. (DNA methylatio...
1. Moreover, the combination of RNA-seq and re-sequencing can be used both to remove false positives when inferring fusion genes and to analyze copy number alterations. 2. The statistically significant correlations that were observed, however, accounted for relatively small effects. (DNA methylat...
基于Illumina高通量测序平台的RNA-Seq技术能够在单核苷酸水平对任意物种的整体转录活动进行检测,在分析转录本的结构和表达水平的同时,还能发现未知转录本和稀有转录本,精确地识别可变剪切位点以及cSNP(编码序列单核苷酸多态性),提供最全面的转录组信息。测序深度(Sequencing Depth)指测序得到的碱基总量与目标测序片段大小的...
fragment:实验中样品 total RNA/DNA 随机打断后产生的片段。 sequencing depth:测序深度。测序得到的总碱基数与待测基因组大小的比值。假设一个基因大小为 2M,测序深度为 10X,那么获得的总数据量为 20M。 Illumina/HiSeq MiSeq:Illumina 二代测序仪顶级提供商,Hiseq 与Miseq 为其两款测序仪。Miseq 相比于 Hise...
RNA-seq数据分析缺少公共lncRNA数据库,无法快速的系统性注释和分析lncRNA 不像蛋白编码基因已具有成熟的参考数据库,RNA-seq目前仍缺少公共的完善可靠的参考数据库,以用于原始测序数据的序列比对和注释。此外,RNA-seq 的短reads 在5’末端或3’末端覆盖度不均一,且经常存在RNA降解、或者逆转录过程不能完整的复制至...
8. Liu, Y., et al., Evaluating the impact of sequencing depth on transcriptome profiling in human adipose. PLoS One, 2013. 8(6): p. e66883. 9. Steijger, T., et al., Assessment of transcript reconstruction methods for RNA-seq. Nat Methods, 2013. 10(12): p. 1177-84. ...