Introduction to Sequencing Platform
Sequencing by synthesis (SBS) technology
Illumina sequencing-by-synthesis (SBS) technology is the most successful and widely used next-generation sequencing platform in the world. Massively parallel sequencing is supported by SBS technology, which utilizes a patented end-terminator-based method to detect individual bases as they are incorporated into an extending DNA strand. The fluorescently labeled terminator is imaged as each dNTP is incorporated and subsequently cleaved to effect incorporation of the next base. Since all four reversible terminator bound dNTPs are present during each sequencing cycle, natural competition minimizes incorporation bias. The end result is true single-base sequencing, which obtains the most accurate data in the industry and is suitable for a wide range of applications.
NovaSeq 6000 system
The NovaSeq 6000 system represents the most powerful, simplest, scalable and reliable high-throughput Illumina sequencing platform to date, producing high-quality data. The instrument uses well-established Illumina sequencing-by-synthesis technology. A reversible terminator-based method can perform massively parallel sequencing of billions of DNA fragments and detect single bases as they are incorporated into growing DNA chains. The method significantly reduces the associated errors caused with strings of repetitive nucleotides (homopolymers). Higher throughput: two Flow Cells per Run can generate not less than 2Tb of data; Shorter cycle: running time is less than 40h, producing 6Tb of data; Wider application: applicable to all species and various library types.
HiSeq 2500 Sequencing System
The HiSeq 2500 System is a powerful and efficient ultra-high throughput sequencing system that supports the widest range of applications and research scales. The HiSeq 2500 system features two operating modes, a fast operating mode and a high production operating mode, capable of handling one or two flow cells simultaneously. The fast run mode provides fast results, can efficiently handle limited samples, and supports double-ended 250 bp read lengths, which allows deeper coverage and improved assembly for de novo applications.
Whole genome and candidate region resequencing, transcriptome analysis, small RNA discovery, methylation profiling and whole genome protein-nucleic acid interaction analysis, among others, we have designed complete solutions to simplify any study you want to conduct. Whatever your research goals, lab size, or funding, the HiSeq 2500 system provides an ideal solution for all your high-throughput sequencing needs.
PacBio Sequel is the third sequencing system launched by Pacific Biosciences company after RS and RSII systems. Compared with RS and RSII, the number of zero-mode waveguides(ZMWs) of a single cell in Sequel has increased from 150,000 to 1,000,000. The data output of a single cell will be about 7 times that of the previous one, but the volume is only 1/3 of Pacbio RS II, the sequencing cost is significantly reduced, and the project cycle is greatly reduced. The core technology of the platform is Single molecule Real-Time (SMRT). The SMRT Cell is used as the carrier for sequencing reaction. The fluorescent signal detection area located at the bottom of the ZMW pore anchors DNA polymerase and a DNA fragment, and captures the signals of different bases through the process of different fluorescently labeled nucleotides and fluorescence excitation, thus obtaining DNA sequence information. PacBio Sequel sequencing has revolutionized the problem of the length of second-generation sequencing reads and the inability to splice high-repeat regions, and is now mature for scientific research and clinical applications.
Platform advantages: ultra-long reading length: the average reading length is 10-15kb, and the longest reading length is more than 60kb; Very low bias: no PCR amplification, no GC preference, uniform coverage; Accuracy: the accuracy of the consistent sequence can reach 99.999; High detection rate.
Pacbio Sequel II
The Sequel II system includes the new 8M SMRT Cell sequencing chip and supporting reagents. The instrument control software and SMRT Link software package have been updated. Compared with the previous Sequel System, Sequel II can significantly reduce project cost and time, and the data yield exceeds the former by eight times. Using the Sequel II system, customers can comprehensively detect variations in the human genome ranging from single nucleotide mutations to structural variations. The Sequel II system is also ideal for de novo assembly of large genomes and for full-length transcriptome analysis using Iso-Seq methods. Extensive analysis of Sequel II data shows that the system can provide the same or higher accuracy than the Sequel System. HiFi sequencing mode can realize comprehensive human variation detection. Under the condition that the genome coverage is only 15 times, using HiFi reads to re-sequence the human genome can obtain ≥ 95% sensitivity and specificity for structural variation, ≥ 99% sensitivity and specificity for single base mutation, and ≥ 96% sensitivity and specificity for small fragment insertion and deletion.
Miseq Sequencing System
Miseq is a personal second-generation sequencer Illumina launched in 2012, which uses Illumina proven TruSeq.™Side-by-side sequencing technology, but the price is much lower. It is the only next-generation sequencer that integrates amplification, sequencing, and data analysis on a single instrument, generating more than 10 Gb of data per run, in a footprint of only 2 square feet. MiSeq's revolutionary processes and unparalleled accuracy make it the ideal platform for fast and cost-effective genetic analysis for a wide range of applications.
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