Poly(A) Site Sequencing (PAS-Seq) is a next-generation sequencing-based method that is extensively used in quantitatively profiling of RNA polyadenylation at the transcriptome level. PAS Sequencing provides valuable information for identifying both poly(A) junctions in mRNAs and noncoding RNAs in an accurate and comprehensive manner, as well as provides quantitative information on the relative abundance of polyadenylated RNAs. PAS-Seq studies of different bio-development processes and diseases provide us with a systematic approach to understand biological regulation beyond the level of gene expression, which is a research direction worth to be further studied.
Overview
The alternative polyadenylation (APA) mechanism generates a distinct 3'-termini on mRNAs and other transcripts. 3'-UTR cis-regulatory elements can affect the regulation of the transcription. The approach of studying APA allows researchers to gain more understanding of miRNA regulation, mRNA stability and localization, and mRNA translation. In addition to 3'-Tag-RNA-Seq method, PAS-Seq provides an alternative method for APA analysis. In this process, mRNA is cleaved into fragments of 150 bp or so. Template replacement is then performed using primers with oligo-dT to generate cDNA for subsequent sequencing.
Features
High Performance
Transcriptome-Wide
Bioinformatics Analysis
One-stop Service
High resolution, sensitivity, and specificity
A systematic approach to understand biological regulation beyond the level of gene expression
Our integrated bioinformatics pipeline can be tailored to suit your project.
Provides one-stop service for library construction, sequencing, sample QC and data analysis.
Project Workflow
1. Sample Preparation
RNA purification; quality assessment and quantification
2. Library Preparation
RNA fragmentation; cDNA library preparation.
3. Sequencing
Illumina HiSeq; PE 50/75/100/150.
4. Data Analysis
Visualize and preprocess results, and perform custom bioinformatics analysis.
Bioinformatics Analysis Pipeline
In-depth data analysis:
Biostatistical analysis - transcript copy number comparisons, expression analysis, length distribution, multi-parameter data analysis, etc.
GO and KEGG enrichment analysis
Detect novel and rare transcripts
Target gene prediction and functional analysis
Alternative polyadenylation
UTR isoform analysis
Sample Requirements
RNA sample quantity ≥ 50 ug.
Please make sure that the RNA is not significantly degraded.
Sample storage: RNA can be dissolved in ethanol 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 sealing film. Shipments are generally recommended to contain 5-10 pounds of dry ice per 24 hours.
Ashar-Patel A, Kaymaz Y, Rajakumar A, et al. FLT1 and transcriptome-wide polyadenylation site (PAS) analysis in preeclampsia. Scientific reports. 2017, 7(1): 1-14.
Yao C, Shi Y. Global and quantitative profiling of polyadenylated RNAs using PAS-seq. Polyadenylation: Springer; 2014. p. 179-85.
Shepard PJ, Choi E-A, Lu J, et al. Complex and dynamic landscape of RNA polyadenylation revealed by PAS-Seq. RNA. 2011, 17(4): 761-72.
* For Research Use Only. Not for use in diagnostic procedures.
