- de novo Sequencing
- Whole Genome Resequencing
- Exome Sequencing
- Target Region Sequencing
- Genotyping by Sequencing
- Whole Genome Mapping
- Sanger Sequencing
- Single-Cell DNA Sequencing
- Human MHC-Seq
- Single-Cell Sequencing
- Immune Repertoire Sequencing
- FFPE Samples
Genotyping by sequencing (GBS) is a unique, cost-effective tool for associate studies and genomics-assisted breeding. It generates large number of single nucleotide polymorphisms (SNPs) for use in genetic analysis. GBS is becoming increasingly important, particularly in plant species with complex genomes that lack reference sequences. We use methylation-sensitive restriction enzymes to reduce genome complexity and avoid the repetitive fraction of the genome, where methylation is more likely to happen, thus allowing lower copy regions to be targeted with two- to three-fold higher efficiency. Other key advantages of this system include reduced sample handling, fewer PCR and purification steps, no size fractionation, and inexpensive bar-coding. Further applications of GBS to breeding, conservation, and global species and population surveys allow plant breeders to conduct genomic selection on a novel germplasm or species without first having to develop any prior molecular tools, and conservation biologists to determine population structure without prior knowledge of the genome or diversity in the species.
- Decreased amounts of input DNA: Only 300 ng of each sample is required.
- Faster, simpler protocol compared to traditional methods: The protocol takes 40 working days to complete for 96 samples.
- Large range of applications: GBS is suitable for species with or without reference sequences.
- Low cost: GBS is an attractive and feasible option for large numbers of markers or individuals.
Library Construction Workflow:
Figure 1. Steps in GBS library construction, including barcoding, enzyme digestion, and PCR amplification.
- Standard bioinformatics analysis
- Basic analysis of sequence data
- Reads clusters across tagged sites of the sequence in species without reference sequences; alignment for species with reference sequences
- SNP detection
- Advanced bioinformatics -- Population analysis
- Phylogenetic tree analysis
- Population structure analysis
- Principal Component Analysis (PCA)
- Advanced bioinformatics -- Genetic map construction
- Genetic map construction
- QTL mapping analysis (phenotype data provided by the customer)
- Integration with the original genetic map for the same mapping populations if provided by the customer
- Sample type: genomic DNA with no or minimal degradation and contamination
- Sample quantity: ≥ 300 ng/sample
- Sample concentration: ≥25 ng/µL
- Sample number: 96 or in multiples of 96