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Request a QuotePlasmid-mediated horizontal gene transfer is considered a major driving force for rapid bacterial adaptation and diversification. Identification of unknown or known plasmids was once difficult due to repeat regions. CD Genomics employs next generation sequencing (NGS) or long-read sequencing technology to accurately obtain all genomic knowledge of plasmid. Coupled with powerful bioinformatics algorithms and tools, we can help you identify antibiotic resistance genes (ARGs), mobile genetic elements and any other genes of interest.
Plasmids are small and circular pieces of DNA presenting in bacteria (like Escherichia coli) and other organisms. It can replicate independently in host genome. Plasmids are important vehicles for microbial populations to adapt rapidly to environmental changes. Plasmid-mediated gene transfer is crucial in the dissemination of ARGs. Plasmids are transmissible between host cells, leading to the quick spread of their accessory genes within a bacterial community. The Food Safety Authority (EFSA or FDA) requires critical plasmid information in food or feed industry, mainly because of the transmission of antibiotic resistance traits.
Whole-plasmid sequencing refers to the technique that utilizes high-throughput sequencing technology to sequence an entire plasmid genome. This process involves library construction of the plasmid genome for sequencing. The sequencing data is subsequently subjected to bioinformatics analyses, wherein aspects such as sequencing depth, GC content, and the assembly of the full plasmid sequence are systematically examined and analyzed.
Sanger Sequencing | Whole Plasmid Sequencing | |
---|---|---|
Data Quality | Peaks, double peaks; requires manual analysis and correction | Superior quality; no need for manual correction |
Sequencing Length | 1 kb | 2.5 - 300 kb |
Cost | Low | Lower |
Sample Quantity | 100 ng | 1-100 ng |
Accuracy | 70-90% | ≥97% |
Sequencing Capability | Requires sequencing primers; limited to sequencing primer-targeted fragment sequences | No need for sequencing primers; capable of sequencing the entire plasmid length |
Throughput | Low | High |
Experimental Timeline | Several tens of days | A few days |
NGS and long-read sequencing technologies offer a strong potential for plasmid identification, diversity, characterization and evolution studies. Based on Illumina HiSeq, PacBio or Oxford nanopore sequencing platforms, we are dedicated to offer full-length plasmid sequencing service, to detect and identify plasmids. Long-read sequencing can span complex repetitive regions to ensure complete and accurate sequence data. We help you discover mobile genetic elements, screen ARGs, annotate other accessory genes. Good annotation of plasmid genomes is necessary to maximize the value of the rapidly increasing volume of bacterial strains. These information are key when determining food safety & quality and controlling for antimicrobial resistance.
At CD Genomics, we offer a comprehensive modular plasmid design approach to meet diverse requirements. Whether you require plasmids with repetitive sequences, high or low GC content, exceptionally long target sequences, or plasmids with multiple transgenes, we can assemble plasmids tailored to your specific needs. Our team of experts brings years of experience in plasmid engineering, ensuring the precision of each designed plasmid. If the desired plasmid type is not readily available, rest assured, as we will closely collaborate with you to design and construct plasmids that align with your exact specifications.
Sequencing Platform | Library Type | Sequencing Depth |
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Illumina 150PE/250PE | 400~500 bp | 100X |
PacBio SMRT | 2 K | 100X |
Our bioinformatics analysis includes data quality control, genome assembly, functional annotation and comparative genomic analysis, which is flexible to your needs. Please feel free to contact us to discuss your project.
Bioinformatics Analysis | Details |
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Data QC | Data resource: Illumina (PE 75/250, 100X) or PacBio (4 Kb library, 100X) |
Read quality assessment, trimming and filtering | |
Sequence assembly | De novo assembly and outcome evaluation |
Functional Annotation | NCBI GenBank, SwissProt, ARDB (Antibiotic Resistance Genes Database), COMBREX, Snapgene, PlasMapper, etc. |
Sample type | Category | Size | Concentration | Minimum volume |
---|---|---|---|---|
Plasmid | Standard | 2.5 -25 kb | 30 ng/uL | ≥10 uL |
Big | 25-125 kb | 50 ng/uL | ≥20 uL | |
Huge | 125-300 kb | 50 ng/uL | ≥40 uL | |
Linear/Amplicon | Standard | 600 bp - 25 kb | 30 ng/uL | ≥10 uL |
Big | 25 - 125 kb | 50 ng/uL | ≥20 uL |
Please feel free to reach out if you have any further inquiries or require additional information.
Reference
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