Lab 06 Notes.docx

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Department
Cell and Systems Biology
Course
CSB352H1
Professor
Ron Ammar
Semester
Winter

Description
Lab 6: Next Generation Sequencing Applications – Summary RNA-Seq (Whole transcriptome sequencing) • The study of an organism or community’s transcribed genes through next-generation sequencing of steady-state RNA from the sample of interest • Quantitative examination of transcriptional profile of an organism or community – what genes are “on” and levels of transcription o Indicates functional activity • RNA reverse transcribed to cDNA prior to sequencing • Double stranded cDNA molecules are sheared into smaller fragments to which adapters are then ligated • Fragments are sequenced o Single end reads o Paired-end reads – short reads from both ends are generated, leading to a better mapping of ambiguous reads • Normalization – allows to compare expression results • Splicing – refers to the removal of intr
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