Transcriptome analysis by strand-specific sequencing of complementary DNA.
High-throughput complementary DNA sequencing (RNA-Seq) is a powerful tool for whole-transcriptome analysis, supplying information about a transcript's expression level and structure. However, it is difficult to determine the polarity of transcripts, and therefore identify which strand is transcribed. Here, we present a simple cDNA sequencing protocol that preserves information about a transcript's direction. Using Saccharomyces cerevisiae and mouse brain transcriptomes as models, we demonstrate that knowing the transcript's orientation allows more accurate determination of the structure and expression of genes. It also helps to identify new genes and enables studying promoter-associated and antisense transcription. The transcriptional landscapes we obtained are available online.
Parkhomchuk
Dmitri
D
Max Planck Institute for Molecular Genetics, Department of Vertebrate Genomics, Ihnestr. 73, 14195 Berlin, Germany.
Borodina
Tatiana
T
Amstislavskiy
Vyacheslav
V
Banaru
Maria
M
Hallen
Linda
L
Krobitsch
Sylvia
S
Lehrach
Hans
H
Soldatov
Alexey
A
D000818:Animals; D018076:DNA, Complementary / Q000737:chemistry*; D003856:Deoxyuracil Nucleotides / Q000378:metabolism; D020869:Gene Expression Profiling; D005800:Genes, Fungal; D051379:Mice; D011401:Promoter Regions, Genetic; D016372:RNA, Antisense / Q000096:biosynthesis; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA / Q000379:methods*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D003856:Deoxyuracil Nucleotides; D016372:RNA, Antisense; C027078:deoxyuridine triphosphate