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DNA/RNA Sequencing – Nanopore Sequencer
ONT sequencing is a single molecule real-time electrical signal sequencing technology based on nanopores, the sequencing principle of each platform is the same. Double-stranded DNA/RNA will bind to nanoporous protein embedded in the biofilm and unwinding under the lead of motor protein, under the action of voltage difference from both sides of the biofilm, DNA/RNA strands pass through the nanopore channel protein at a certain rate. Due to the differences of the chemical properties of the different bases on the DNA/RNA strand, when a single base or DNA molecule passes through the nanopore channel, it will cause the change of different electrical signals. By detecting and corresponding to these signals, the corresponding base types can be calculated, and the real-time detection of the sequence can be completed.
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DNA/RNA Sequencing – Illumina Sequencer
Illumina sequencing technology, sequencing by synthesis (SBS), is a widely adopted next-generation sequencing (NGS) technology worldwide, responsible for generating more than 90% of the world’s sequencing data. A fluorescently labeled reversible terminator is imaged as each dNTP is added, and then cleaved to allow incorporation of the next base. Since all 4 reversible terminator-bound dNTPs are present during each sequencing cycle, natural competition minimizes incorporation bias. The method virtually eliminates errors and missed calls associated with strings of repeated nucleotides (homopolymers). Illumina sequencing by synthesis technology supports both single-read and paired-end libraries. SBS technology offers a short-insert paired-end capability for high-resolution genome sequencing, as well as long-insert paired-end reads for efficient sequence assembly, de novo sequencing, and more. The combination of short inserts and longer reads increases the ability to fully characterize any genome.
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DNA/RNA sequencing -PacBio Sequencer
PacBio sequencing platform is a long-read sequencing platform, which is also known as one of the Third-Generation Sequencing(TGS) technologies. The core technology, single-molecule real-time(SMRT), empowers the generation of reads with tens of kilo-base in length. On base of “Sequencing-by-Synthesis”, single nucleotide resolution is achieved by Zero-mode waveguide(ZMW), where only limited volume at the bottom(site of molecule synthesis), is illuminated. In addition, SMRT sequencing largely avoids sequence-specific bias in NGS system, in that most of PCR amplification steps are not required in library construction process.
Platform: Sequel II, Revio
