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| Next-generation sequencing technology |
Next-generation sequencing is a technique that allows ultra-high throughput speed to analyze DNA. There are two main types of next-generation sequencing, one with a short read and the other with a long read. In the future next-generation sequencing technology can be how they are implemented for disease diagnoses such as cancer and covid-19.
Next-Generation Sequencing
Next-generation sequencing is a massive technique that allows ultra-high throughput speed to analyze DNA. The method is used to find nucleotides of the whole genome or specific DNA portion. Next-generation sequencing technology has changed biology, allowing labs to undertake a wide range of applications and analyze living mechanisms with more feasibility.
Next-Generation Sequencing Techniques:
Illumina
sequencing: This
technique recognizes DNA base pairs and adds them to a nucleic acid chain. Each
base emits a distinct fluoresce cent signal.
Pyro
sequencing: The
technique of pyrosequencing known as Roche 454 sequencing, is an approach that detects
phosphate release using a light signal after nucleotides are incorporated by the polymerase into the new strand.
Proton sequencing/Ion Torrent: Ion Torrent sequencing detects the emission of a proton from a cell.
Advantages of NGS
Next-generation sequencing is a common technology in functional genomics
for analyzing DNA and RNA.
- ·
It has a single-nucleotide resolution,
allowing it to identify associated genes, gene alleles, gene variants, and
single nucleotide polymorphisms without having knowledge of genomic features.
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Requires less DNA/RNA as input data
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Has a higher dynamic range of signals
- · More repeatability
Types of Next-generation Sequencing:
There are basically two types of sequencing reads: one with small, precise
reads known as Ion Torrent and the other with long runs and reads known as Nanopore.
Sequencing of short-reads:
Illumina and Ion Torrent is a short-read method with a limited read size of 400 bases. It can normally sequence anything but repetitive and duplicated genomic areas offer a challenge because it is difficult to discover unique sections of the target gene to compare to the control sample.
The Human Leukocyte Antigen system, which codes for components of immunological function, is one example of a therapeutically important gene that can be sequenced by this technique. It’s hard to put together in small pieces, but it's much easier in large pieces.
Single DNA molecules are immobilized in small wells using optical technology. Each DNA polymerase will be separately monitored and all the copies will be amplified. A fluorescent label is added to the end of a phosphate group. During synthesis, when the polymerase chain cleaves the phosphodiester bond of DNA, it gives fluorescence upon emission. Longer DNA segments consist of up to large base pairs that can be read by new-generation sequencers. Up till now, these small read techniques had a reputation for being prone to errors.
Sequencing of Long-reads:
The other main long-read technology is Oxford
Nanopore, which can read up to 4 million bases. The
electric-resistant membrane trapped a single DNA molecule. The current is
disturbed as the DNA molecule passes through nanopore embedded electric membrane. DNA or RNA sequences of molecules can be
analyzed by real-time PCR. This means that sequencing is substantially faster,
and single molecules can be sequenced in a couple of minutes. Nanopore
Technology produced its first portable pocket-sized sequencing equipment in
2014. It can create lengthy runs.
Hundreds of gaps remained in areas of structural variation including repeated missing spaces that may be associated with disease when the human genome was sequenced in 2001. The Telomere-to-Telomere (T2T) group completed the whole genomic sequence although it was imperceptible to Illumina short-read sequencing technique.
F Future of Next-generation Sequencing:
Even though next-generation sequencing has made an evolution in the field of Science. We can apply Next-generation sequencing for the diagnosis of diseases. Now we can move forward and apply these techniques to benefit human health.
Cancer Diagnostics:
The future of Next Generation Sequencing firms
is in creating new applications. The existence of tiny DNA fragments in the blood
is one of the breakthroughs that have opened the way. Companies have now gone
on to cancer diagnosis, having first utilized it for non-invasive DNA testing
of fetal DNA.
The Illumina technology delivers information on
mutations that occurred in around 500 genes discovered in tumors. The challenge is
to create a next-generation sequencing-based diagnosis. This method is also being developed by
Ion Torrent. Next-generation sequencing makes it simple both from a technical
and bioinformatics point of view.
Next-generation sequencing techniques in the future will cure infectious diseases including cancer. Some healthcare systems are beginning to embrace human genome sequencing, which seeks to sequence cancer and rare illness in patients.
The Covid-19 challenge in infectious disease surveillance:
Illumina created monitoring kits for COVID-19, which
served as a baseline against other viral genotypes that could be analyzed. This
is important because it allows us to create a surveillance program for
diseases.
Ion Torrent technology has been expanded to provide the survival of diseases. It boosted immunity. With this technology in the future, the whole genome can be sequenced within 20 viral copies which are very low so the viral load is lower. The LamPORE assay is a low-cost technology that is highly scalable for COVID-19 screening tests. This technique identifies viruses in just a few minutes and was created by Oxford Nanopore. LamPORE sequences RNA and amplifies conserved genes that are present in COVID-19.
Conclusion:
In the next years, it's quite likely that you'll get a blood-based cancer test that can detect the early stages of cancer and will frequently be used to maintain your health. Next-generation sequencing is also used for the diagnosis of other disorders.
In the future, genome sequencing may be included in routine medical exams, allowing mutations and epigenetic alterations to be followed over time. However, we may have to wait a bit longer for these advancements until our ability to comprehend the human genome catches up to the information that sequencing technology can now diagnose diseases.
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| Next-generation sequencing technology for disease diagnosis |
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