SMRT full-length RNA-sequencing is a third generation sequencing (TGS)-based method to amplify and sequence the whole transcriptome of cells or tissues. We provide SMRT full-length RNA-sequencing to help you identify new transcripts and alternative splicing events, resolve structural variants and repetitive regions.
SMRT full-length RNA-sequencing is one kind of long-read sequencing, emerged in 2011, provided by Pacific Biosciences (PacBio). SMRT is short of single molecule real-time, thus allowing you to distinguish alternative transcript isoforms at the single-cell level. SMRT full-length RNA-sequencing based on sequencing by synthesis. The RNA sample was reverse-transcription to cDNA, as DNA polymerase incorporated labeled dNTPs into the single-stranded cDNA template, the fluorescence pulses were recorded. There are two sequencing modes: circular consensus sequencing (CCS) and continuous long read (CLR) sequencing. The CCS mode is more commonly used, with highly accuracy, the average read is 13.5 kb. CLR mode allows >50 kb reads. Advantages of SMRT full-length RNA-sequencing include: 1) no assembly required with less errors in repetitive sequences; 2) capture cell-to-cell heterogeneity and regions between alleles; 3) deliver a more complete view of genetic diversity.
We provide SMRT full-length RNA-sequencing services for identifying new genes, transcripts, and alternative splicing events, resolving structural variants and repetitive regions. SMRT full-length RNA-sequencing increases the accuracy of RNA-seq at the isoform-level resolution, it allows a high--resolution view of full-length cDNA sequences without assembly.
|High throughput||Single-Cell Insights||High Resolution||Multiple Applications|
|This method allow long reads for accurate analysis with a short sequencing time.||Quantitative analysis down to single-cell levels for input samples.||Discovery of more cellular differences based on high resolution analysis.||Identify new genes, transcripts and alternative splicing events.|
RNA purification; quality assessment and quantification.
Single-cell cDNA enrichment; cDNA library preparation.
Circular consensus sequencing (CCS); continuous long read (CLR) sequencing.
Visualize and preprocess results, and perform custom bioinformatics analysis.
RNA sample (quantity ≥ 100 pg), 1.8 ≤ OD260/280 ≤ 2.2, OD260/230≥2.0, RIN ≥ 6.5, 28S:18S≥1.0. Please make sure that RNA is not significantly degraded.
Sample Storage: The sample should be stored at -80°C. Avoid repeated freezing and thawing.
Shipping Method: When shipping the sample, it is stored in a 1.5 mL Eppendorf tube, sealed with a sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
Deliverable: FastQ, raw data, coverage summary, QC report, experiment results, custom bioinformatics analysis.