The best RNA-seq workflow for your experiment depends on the questions you are trying to answer, the quality of the RNA sample, the amount of RNA available, and the nature of the transcripts of interest. The KAPA RNA HyperPrep Kit is a robust, streamlined library preparation solution for the preparation of total RNA libraries, and forms the basis for KAPA RNA-seq workflows for transcriptome sequencing with ribodepletion, custom depletion, and mRNA capture workflows (see below).
Depletion of rRNA, user-defined RNA, and globin RNA
Whole-transcriptome sequencing provides a complete view of the transcriptome, including immature and noncoding RNAs. In these workflows, it is important to remove rRNA (up to 95% of the total RNA sample) prior to library prep in order to minimize the number of wasted sequencing reads. This method is effective with degraded RNA inputs not suitable for mRNA capture. Enzymatic depletion can also be used to remove other transcripts, such globin mRNA and/or other highly abundant RNAs.
mRNA capture sequencing is used for the analysis of coding transcripts in eukaryotic samples, allowing researchers to detect and quantify relative expression levels. This workflow enriches for mRNA over non-polyadenylated species such as ribosomal, precursor, and non-coding RNAs.
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