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CD Genomics is now providing 2b-RAD sequencing which is a novel reduced-representation whole genome sequencing and restriction-site associated DNA (RAD) sequencing method for linkage mapping and determination of genome-wide variants in a cost-effective way.

Introduction to 2b-RAD

2b-RAD method uses type IIB restriction enzymes (REs), which share the feature of cutting the genomic DNA at both sides of the recognition to produce a fixed-size dsDNA fragment and result in protruding, noncohesive ends. Subsequently, DNA fragments of interest are captured by a biotinylated adaptor specific to the initial enzyme. Compared with 2b-RAD, similar methods like restriction-site associated DNA (RAD) and genotyping-by-sequencing (GBS) technologies lack the specificity to regions of interest, and produce many sequences originated from non-informative and repetitive regions.

Sequencing of these short and uniform tags brings many benefits, including targeting of all restriction sites, even sequencing depth across sites, high reproducibility for quantitative measurement, low sequencing cost per tag and low sensitivity to DNA degradation. To benefit from the gradually increased sequencing capacity of next-generation sequencing (NGS) platforms, we also combine an advanced protocol that allows the preparation of five concatenated isoRAD tags for Illumina paired-end (PE) 150 bp sequencing, which provides researchers more power and flexibility in devising effective library configurations to meet specific research purposes.


  • Bin Map construction and QTL location
  • Population genetic study
  • Population evolution analysis
  • Genome-wide association study
  • Genome phylogeny
  • Genomic selection
  • Linkage and association mapping
  • Discrimination of microbial strains
  • Detection of somatic mutations

Advantages of 2b-RAD

  • Accurate and affordable
  • Flexible tag number
  • Consistent label length
  • High density of markers
  • Not requiring interim purification steps
  • Allowing low DNA input and degraded DNA
  • Highly reduced 2b-RAD libraries require much less sequencing for accurate genotyping

2b-RAD Workflow
The general workflow for 2b-RAD includes sample preparation, 2b-RAD library preparation, high-throughput sequencing and bioinformatics analysis. Our highly experienced expert team executes quality management, following every procedure to ensure confident and unbiased results. The library construction for 2b-RAD consists of four major stages (BsaXI digestion, ligation, amplification and barcoding).

Service Specifications

Sample Requirements

  • DNA amount ≥ 100 ng; DNA concentration ≥ 20 ng/µl; OD 260/280 = 1.8~2.0
  • Species: Diploid organisms with or without a reference genome
  • All DNA samples are validated on purity and quantity

  • Illumina HiSeq X ten, PE 150
  • Genome label average density: 2 kb
  • The average depth of label ≥ 20X
  • Classification accuracy ≥ 95%
Bioinformatics Analysis

  • Raw data quality control
  • Statistical analysis
  • SNP genotyping
  • Annotation of SNP locus
  • Construction of genetic linkage map
  • Population studies

CD Genomics is proud to offer advanced 2b-RAD sequencing technology. Our service covers the complete process, from quality control of genomic DNA to determination of genotypes and population diversity. If you have additional requirements or questions, please feel free to contact us. CD Genomics is the licensed service provider for this technology. Keygene N.V. owns patents and patent applications protecting its Sequence Based Genotyping technologies.

* For Research Use Only. Not for use in diagnostic procedures.
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CD Genomics
CD Genomics-the genomics service company