Principle of CAPS and dCAPS Markers

RNA sequencing or Whole Transcriptome Shotgun Sequencing is next generation sequencing method provide snapshot of coding and non-coding transcripts to understand connection between genome and its gene expression and identify novel features of gene fusions and isoforms, Direct RNA sequencing perform by sequence isolated RNA directly without converting to cDNA while in-direct RNA sequencing or complementary RNA sequencing perform by sequence cDNA after converting from isolated RNA, Whole transcription RNA sequencing or Total RNA sequencing sequence all types of RNAs to target both coding and non-coding transcripts to provide all information about gene expression, mRNA sequencing sequence Poly-A library (directly or in-directly) to identify expressed coding genes, Target RNA sequencing sequence specific transcript to study particular genes or pathways, Small RNA sequencing sequence microRNA and short interfering RNA and Piwi interacting RNA to study gene regulation, Single cell RNA sequencing sequence RNA from individual cells to characterize cellular heterogeneity within complex cell and identify unknown and rare cell types and explore changes in gene expression among cell type and trace trajectory of distinct cell lineage, RNA Sequencing initiate by RNA extraction and RNA selection to select target RNA and synthesize cDNA from isolated RNA and prepare cDNA library to generate millions reads of each sample in process of fragmentation to optimize sequencing of target cDNA and adaptor ligation to bind transcripts as priming site for sequencing and index and amplification by add specific sequence or barcode to transcripts during PCR amplification of cDNA to increase concentration of developed cDNA library for sequencing by next generation sequencing and analyze obtain data by bioinformatics tools by comparing obtain sequence with reference sequence, RNA Sequencing detect differences in gene expression levels without prior knowledge of genome sequence to detect novel transcripts and detect allele specific expression and detect alternative gene splicing transcripts and detect post transcriptional modifications and detect gene fusions and isoforms and other structural variants, RNA Sequencing consider labor and costly and time consuming method because analyzed RNA sequencing data require significant computational resources and experts, RNA Sequencing use in mutation studies and gene expression studies and disease diagnosis studies and biomarker studies and epigenetic studies and functional genomic studies and Pharmacogenomics studies, Systems of RNA sequencing (I-sequence 100 system and next sequence 1000 and next sequence 2000 and nova sequence 6000 system and nova sequence X series) (Watch Related Video in #geneticteacher) #geneticteacher