Label-free DIA Quantitation

Service Description

BGI Genomics participates in quantitative performance assessment as well as standardization and harmonization of Multi-National DIA proteomics analysis supporting precision medicine studies. We have developed innovative solutions for optimizing the process of accurately quantifying proteins in complex biological matrices and other complex mixtures.

Sample Preparation and Services

 

 

  • The QC analysis of enzymatic hydrolysis is strictly controlled by LC-MS
  • DDA-Spectral Library generated using HPLC technology and sample fractionation
  • 2h/fraction for non blood; and 2h/fraction for non-human blood; 1h/fraction for human blood

Quality Standard

 

  • Summary includes all methods and data analysis
  • Reports provided in Excel or PDF format, RAW files available upon reques

Turn Around Time

 

  • Typical 4-5 weeks from sample QC acceptance to data report delivery

Label-Free DIA Quantitative Proteomics

Data-Independent Acquisition (DIA) is a label-free quantitative technique which provides highly consistent quantitation and broad proteome coverage. We create a customized spectral library with your specific sample and then analyze individual samples using nano-flow LC-MS/MS with DIA scanning and quantitation.

Project Workflow

Sample preparation

HPLC FRACTIONATION

LC-MS/MS DDA Analysis

DDA library generation and updates

LC-MS/MS DIA analysis

Quantitation and bioinformatics

Specifications

  • The QC analysis of enzymatic hydrolysis is strictly controlled by LC-MS
  • DDA-Spectral Library generated using HPLC technology and sample fractionation
  • 2h/fraction for non blood; and 2h/fraction for non-human blood; 1h/fraction for human blood
  • Summary includes all methods and data analysis
  • Reports provided in Excel or PDF format, RAW files available upon request

Sample Requirements

We accept protein samples in a variety of formats. For attaining maximum proteome coverage we recommend utilizing our sample fractionation services, performed using our off-line UHPLC-UV platform.
Sample type Amount and Concentration Minimum sample volume
Protein sample in liquid solution Recommended 400 µg with fractionation: 2 µg/µL 200 µL
Protein sample in liquid solution Minimum Required 100 µg with fractionation: 1 µg/µL 100 µL

 

Advanced Bioinformatics Analysis

  • Data analysis and validation performed with Mascot
  • Protein identification and quantification list
  • Differential proteins data statistics and volcano plot
  • Principal component analysis (PCA) and cluster analysis
  • Protein GO/COG/KOG/Pathway annotation
  • GO/COG/KOG enrichment analysis of differential proteins
  • Protein-protein interaction analysis
  • Protein subcellular localization analysis
  • Time series analysis
  • Repeatability analysis
  • Quantitative proteomics and phosphoproteomics correlation anal

Label-free DIA Quantitation


Unique DNBSEQ™ Sequencing Technology

BGI’s RNA Sequencing services are typically executed with proprietary DNBSEQ™ sequencing technology platforms, for great sequencing data at some of the lowest costs in the industry. DNBSEQ™ offers advantages in terms of lower amplification error rates and much lower duplication rates. In addition, studies have shown the lower index hopping rate in DNBSEQ™ platforms.