High-Throughput Sequencing for Trypanosome Transcriptome Characterization

Methods Mol Biol. 2020:2116:83-98. doi: 10.1007/978-1-0716-0294-2_6.

Abstract

High-throughput sequencing of cDNA (RNASeq) is now the method of choice for analysis of transcriptomes. This chapter details important considerations in the design of RNASeq experiments for kinetoplastids grown in culture or experimental animals. It contains protocols for obtaining parasites from rodents, and for removal of rRNA from total RNA. In addition, custom pipelines for sequence alignment, and for data analysis and visualization, are described.

Keywords: Gene expression; Gene regulation; Mouse; Trypanosoma; RNASeq; Rat; Rodent; Transcriptome; rRNA depletion.

MeSH terms

  • Animals
  • Disease Models, Animal
  • Humans
  • Mice
  • RNA, Messenger / genetics
  • RNA, Messenger / isolation & purification
  • RNA, Protozoan / genetics
  • RNA, Protozoan / isolation & purification*
  • RNA, Ribosomal / isolation & purification
  • RNA-Seq*
  • Rats
  • Sequence Alignment
  • Transcriptome / genetics*
  • Trypanosoma / genetics*
  • Trypanosoma / isolation & purification
  • Trypanosomiasis / blood
  • Trypanosomiasis / cerebrospinal fluid
  • Trypanosomiasis / parasitology*

Substances

  • RNA, Messenger
  • RNA, Protozoan
  • RNA, Ribosomal