Babak Tomas
Department of Biology, Stanford University, Stanford, CA, USA,
Methods Mol Biol. 2012;925:79-88. doi: 10.1007/978-1-62703-011-3_6.
Enabled by high-throughput technologies that are capable of generating millions of sequencing reads, transcriptome sequencing is emerging as an important approach for mapping allelic imbalance (AI), where transcription is biased toward one allele in a diploid system. AI is identified by counting sequencing reads that map to genomic regions containing heterozygous SNPs, where the base identity of the SNP is used to distinguish allelic origin. Genomic imprinting is a special case of AI where bias is toward parental sex and can be identified by transcriptome sequencing of systems that represent reciprocally inherited loci. The focus of this protocol is on experimental design, analysis, and interpretation of genomic imprint discovery using whole transcriptome sequencing.
借助能够生成数百万条测序读数的高通量技术,转录组测序正成为绘制等位基因不平衡(AI)图谱的一种重要方法,在二倍体系统中,转录偏向于一个等位基因。通过对映射到包含杂合单核苷酸多态性(SNP)的基因组区域的测序读数进行计数来识别AI,其中SNP的碱基身份用于区分等位基因起源。基因组印记是AI的一种特殊情况,其中偏向于亲本性别,可以通过对代表相互遗传位点的系统进行转录组测序来识别。本方案的重点是使用全转录组测序进行基因组印记发现的实验设计、分析和解释。