Ribo-Seq Analysis (Ribosome Footprinting) / New York

Descripción

Ribo-Seq, also known as ribosome profiling or ribosome footprinting, is an advanced transcriptomic research technique based on next-generation sequencing (NGS). It offers a comprehensive and quantitative measurement of translation processes.

Ribo-Seq precisely identifies ribosome positions, revealing actively translated mRNAs. This method enables profiling of active ribosomes within a cell at a specific moment. It also uncovers translation initiation sites, translated ORFs, ribosome locations on mRNAs, and ribosome translation rates.

Ribo-Seq (Ribosome Footprinting)

Ribo-Seq, or ribosome profiling, is an NGS-based technique for comprehensive and quantitative translation analysis. It pinpoints ribosome positions, providing insights into active translation of mRNAs. This method profiles active ribosomes within cells, identifying translation initiation sites, translated ORFs, ribosome-mRNA interactions, and translation rates.

Features

Experience the following benefits with Ribo-Seq:

• High Throughput: Obtain over 5 G information in a single sequencing run.
• High Accuracy: Benefit from accurate reference libraries and extensive sequence coverage, enabling indirect protein identification.
• High Repeatability: Deep sequencing ensures randomness in detection without requiring technical replication.
• One-stop Service: Enjoy a comprehensive service package, covering library construction, sequencing, sample QC, and data analysis.

Project Workflow

1. Sample Preparation: Purification and quantification of RNA, ensuring quality.
2. Library Preparation: Creation of strand-specific cDNA libraries with size selection.
3. Sequencing: Utilization of Illumina HiSeq; PE 150 sequencing.
4. Data Analysis: Visualize and preprocess results, perform custom bioinformatics analysis.

Bioinformatics Analysis Pipeline

Explore a thorough analysis pipeline:

• Data quality control
• Reference-based mapping
• Evaluation of gene expression levels
• Clustering
• GO and KEGG pathway analysis
• Differential gene analysis (translational level)
• Translation ratio analysis
• Protein-protein interaction network analysis

Sample Requirements

For Ribosome Protected Fragment (RPF) samples:

• Concentration: ≥ 200 ng/µL
• Amount: ≥ 2 µg
• Volume: ≥ 10 µL
• Main Peak: 25 – 38 nt

Ensure RNA integrity and purity. We provide Ribosome Nascent-chain Complex (RNC) separation and mRNA extraction services for tissue or cell samples.

Sample Storage

Store RNA in RNA Stable or RNA-free ultra-pure water at -80°C. Avoid repeated freezing and thawing to maintain RNA quality.

Shipping Method

When shipping RNA samples:

• Store RNA in a 1.5 mL Eppendorf tube sealed with film.
• Include 5-10 pounds of dry ice per 24 hours for shipment.

Deliverable

Receive the following upon completion:

• FastQ, BAM
• Coverage summary
• QC report
• Custom bioinformatics analysis

References

Explore these references for further insights:

1. Wang H, Wang Y, Xie Z. Computational resources for ribosome profiling: from database to Web server and software. Briefings in Bioinformatics. 2017, 20.
2. Popa A, Lebrigand K, et al. RiboProfiling: a Bioconductor package for standard Ribo-seq pipeline processing. F1000 Research. 2016, 5.
3. Ingolia NT, Ghaemmaghami S, Newman JR, et al. Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling. Science. 2009, 324(5924): 218-23.

For Research Use Only. Not for use in diagnostic procedures.

Ribo Seq Analysis

For more information:

https://rna.cd-genomics.com/ribo-seq-ribosome-footprinting.html »

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