Compared with conventional approaches like PCR, Whole Genome Sequencing does not require labor-intensive cloning and mapping steps. Hence, it is time- and cost-effective. Moreover, this high-throughput sequencing approach allows the sequencing of numerous samples at the same time through the courtesy of multiplexing.
Applications
- Permits the detection of variations within target genomes
- Interpretation of character differences
- Allows large-scale evolution research
- Enables prerequisite study of novel species identification
Benefits
- Extensive experience: We have successfully completed high-profile projects which cover a wide range of fields such as pathogenic bacteria, probiotics, edible bacteria, medicinal strains, and industrial strains.
- Professional services: From material selection, library construction, and sequencing to data analysis, each step provides scientific and meticulous design to ensure high-quality research results.
- Comprehensive analysis: Detection of SNP, InDel, SV and other mutation information of strain reference genomes, and further research on species evolution, population characteristics, selection pressure, etc. One-stop analysis of variation and difference.
- Strict quality control: High-quality of the sequencing data is ensured by verifying the samples.
- High-quality library preparation: To ensure the quality of the data, all libraries are size selected to optimize the size of the insert.
Specifications: DNA Sample Requirements
| Library Type | Sample Type | Amount (Qubit) | Volume | Concentration | Purity (NanoDropTM/ Agarose Gel) |
| Microbial whole genome library (350bp) | Genomic DNA | ≥200 ng | ≥20 μL | ≥10 ng/μL | A260/280=1.8-2.0; no degradation, no contamination |
| Microbial whole genome library (PCR-free 350bp) |
Genomic DNA | ≥1.2 μg | ≥20 μL | ≥10 ng/μL |
Specifications: Sequencing Parameters and Analysis Contents
| Platform Type | Illumina NovaSeq 6000 |
| Read Length | Paired-end 150 bp |
| Recommended Sequencing Depth | ≥ 100x for bacterial genomes |
| ≥ 50x for fungal genomes | |
| Standard Data Analysis | Data quality control: filtering reads containing adapter or with low quality
Alignment with the reference genome, statistics of sequencing depth and coverage SNP/InDel calling, annotation and statistics CNV calling, annotation and statistics SV calling, annotation and statistics |
Project Workflow
The first step of the project workflow involves the sample quality control (Sample QC) to ensure that your samples meet the criteria of the Microbial WGS technique. Then, the appropriate library is prepared according to your target organism and subsequently tested for its quality (Library QC). Next, a paired-end 150 bp sequencing strategy is used to sequence the samples and the resulting data go through quality data control (Data QC) to guarantee the quality of the resulting data. Finally, bioinformatics analyses are performed and publication-ready results are provided. The following flowsheet describes the step-by-step protocol our Microbial WGS technique follows.
Preparation of sample is followed by the DNA library preparation which is verified for quality and yield. Genomic DNA is fragmented with a size of 350 bp which is narrowly size selected by sample purification beads. The selected fragments are then end polished, A-tailed, and ligated with the full-length adapter. Illumina PE150 technology is employed to sequence the sample and the final stage involves the bioinformatics analysis.
