RNA-Seq

A high-resolution tool for studying changes in the coding and noncoding regions of the transcriptome. RNA-Seq enables the sensitive and accurate measurement of differential gene expression over time or in different groups, conditions and treatments. RNA-Seq also enables the detection of transcript isoforms, gene fusions, single nucleotide variants (SNVs) and allele-specific gene expression.

In principal, RNA molecules are reverse transcribed and tagged with specific “barcoded” oligo adapters, into cDNA fragment libraries. Several cDNA libraries are usually pooled (multiplexing) and sequenced in a single assay. A read (or 2 reads in paired-end) is produced for every cDNA fragment and the total number of reads for each sample are compiled ant then aligned against a reference sequence.

Several RNA-Seq applications can be carried out in GGC, each requiring different types of library preparation and sequencing modes. For differential gene expression (the most common RNASeq application) single-end sequencing with short reads (75bp) and depth ~30M reads is recommended. For other applications, paired-end sequencing and/or higher depth of read may be required.

Total RNA-Seq

Accurately measures gene and transcript abundance and detects both known and novel features in coding and noncoding RNA. Total RNA-Seq can be carried out in both stranded and non-stranded form. Their difference is that in stranded form, the cDNA library is constructed in a way that allows the determination of which genomic DNA strand was transcribed into the RNA.

mRNA-Seq

Accurately measures gene and transcript abundance and detects both known and novel transcript isoforms, gene fusions, and other features as well as allele-specific expression in the coding region of the genome. mRNA-Seq can be carried out in both stranded and non-stranded form. Their difference is that in stranded form, the cDNA library is constructed in a way that allows the determination of which genomic DNA strand was transcribed into the RNA.

Small RNA Seq

Isolates and sequences small RNA species, such as microRNA and other non-coding RNAs, to understand the role of noncoding RNA in gene silencing and post-transcriptional regulation of gene expression.

Single-Cell RNA-Seq

For the study of processes such as differentiation, proliferation, and tumorigenesis, single-cell RNASeq can examine the signals and behavior of a cell in the context of its surrounding environment.