Total RNA-Seq

Total RNA sequencing (RNA-seq) is a next-generation sequencing (NGS)-based method to capture a comprehensive view of the transcriptome. We provide total RNA-seq to help you analyze coding RNAs and multiple forms of ncRNA, including long noncoding RNA (lncRNA), small nuclear (snRNA), small nucleolar (snoRNA), and other ncRNA species.

Overview

RNA transcribed from DNA, is a single-stranded ribonucleic acid molecule and it plays a critical role in coding, decoding, expression and regulation of genes. There are many types of RNA, including mRNA, lncRNA, snRNA, snoRNA and other RNA species. In addition to mRNA, there are many types of non-coding RNAs (ncRNAs) that cannot be translated into proteins and function at the RNA level not protein level. Ribo-Zero ribosomal RNA reduction chemistry can maximize the percentage of uniquely mapped reads and minimize ribosomal contamination. Total RNA-Seq enables the analysis of both mRNA and a broad range of ncRNA species of interest combined the next-generation sequencing and proven ribosomal reduction library preparation chemistries, particularly suited for species whose mRNA may lack a poly-A tail, such as bacteria. It accurately measures gene and transcript abundance, and identify known and novel features of the transcriptome. We provide total RNA-Seq to discover the known and novel features, such as gene fusions, allele-specific expression, alternative transcripts, and the detection of novel transcripts in both coding and ncRNA species.

Project Workflow

Bioinformatics Analysis Pipeline

In-depth data analysis:

• Profiling of lncRNA, snRNAs, snoRNAs and mRNA
• Prediction of novel transcripts
• Target gene prediction and functional analysis of all mapped reads
• GO and KEGG analysis
• Enrichment analysis
• Statistics and annotation of Indels/SNPs
• Biostatistical analysis – expression analysis, alternative splicing analysis, length distribution, multi-parameter data analysis, transcript copy number comparisons, etc.

Sample Requirements

• RNA sample (concentration ≥ 20 ng/uL, amount ≥ 2 ug)
• 1.8 ≤ OD_260/280 ≤ 2.0, RIN ≥ 7.0, 28S/18S ≥ 1.0.
• Please make sure that the RNA is not degraded.

Sample storage: RNA can be dissolved in RNA Stable or RNA-free ultra-pure water and stored at -80°C. RNA should avoid repeated freezing and thawing.

Shipping Method: When shipping RNA samples, the RNA sample 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, BAM, coverage summary, QC report, custom bioinformatics analysis.

Reference:

1. Chang C, Farmer A, Schroth G, et al. cDNA Library Generation for Transcriptome Analysis From Total RNA Equivalent to a Single Cell. J Biomol Tech, 2013, 24: S43.
2. Hayashi T, Ozaki H, Sasagawa Y, et al. Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing and enhancer RNAs. Nature Communications, 2018, 16(5): 239-251.