RNA-Seq for transcriptome analysis

Avance Biosciences offers mRNA-Seq service using Illumina next generation sequencing platform to identify all mRNA transcripts expressed in a biological sample and precisely measure the relative quantities of each transcript including each isoform having different splicing patterns.

Method Description

Biological samples (tissues, blood, etc) preserved for RNA extraction and total RNAs (Rink# >7, 3 μg) extracted by the customer can be submitted for analysis. We will not accept extracted mRNA samples due to the difficulty in assessing their qualities. Sample species should be well characterized with reasonably complete genome information as well as transcript annotation.
The Illumina next generation sequencing platform and its associated mRNA-Seq protocols will be used for analysis.  In summary, messenger RNA with polyA tails will be purified from the total RNA samples. The mRNA will be fragmented and transcribed into ds cDNA for library preparation. Sequencing of the library will be done in a paired-end mode with a read length of 75 cycles.  The paired-end reads will be mapped to the genome, and the potential splicing pattern will be analyzed.  When applicable, expression levels of transcripts, genes, CDSs will be estimated, in RPKM, and reported.

Deliverable

  • Extraction of raw sequencing data (in FASTA or FASTQ format)
  • Alignment of sequencing reads to genome
  • Report of splice junctions
  • Annotation of known transcripts
  • Report of potential new transcripts
  • Report of relative expression levels for transcripts and genes
Contact our customer service for more information about this service and obtain a no-obligation quote.