Sequencing the full complement of mature mRNA
|Platform||Illumina MiSeq, Ion Proton, Ion PGM™|
|Research Objectives||This application can be used for de-novo transcript assembly, identification of genes with low levels of expression, isoform detection, quantification of differential expression in different organisms or different tissues in the same organism, SNP discovery, genotyping, RNA editing, and allele-specific expression|
mRNA-seq allows you to determine steady state mRNA abundance levels by sequencing of full-length poly-A captured transcripts. Inferences regarding co-regulation and the up or down-regulation of genes in tissue specificty and disease states can be made from these data, and allow the detection of dysregulation, alternative transcript isoforms and splicing aberrations.
CPGR will accept purified RNA samples that conform to the minimum requirements. RNA samples will undergo QC using the Bioanalyser. Library preparation and construction will be performed according to project specific requirements followed by high throughput sequencing. The raw FASTQ files are used as an input for standard bioinformatics analysis.
The client will in the first instance receive a sample QC report detailing whether the integrity of the sample meets the requirements for the application. Following sequencing the client will receive an analytical report detailing the parameters of as well as the results of the sequencing run including the raw fastQ files which can be used as the input for standard bioinformatics.
- RNA Quantity:
≥ 5 µg (prokaryotic, human, rat and mouse)
≥ 10 µg (non-model animals)
≥ 20 µg (plants and fungal)
- RNA Concentration:
60 ng/µl (prokaryotic, human, rat and mouse)
150 ng/µl (non-model animals)
250 ng/µl (plants and fungal)