DNA sequencing technique is actually the process for the determination of the nucleic acid sequence and the order type of nucleotides in DNA. It basically includes any technology and method which is used to examine the order of the four bases, which are adenine, guanine, cytosine and thymine.
For quite a long time, Sanger’s chain end strategy was equivalent to DNA sequencing. Two new ages of sequencing stages were dispatched in the mid-2000s which changed the field of transformative genomics.
The primary monetarily accessible cutting edge sequencing strategy was 454 pyrosequencing. This strategy depends on emulsion PCR, and sequencing is accomplished by distinguishing light signals set off by consistently streamed nucleotides. Illumina’s converse eliminator sequencing is the prevailing method utilized for genome sequencing today.
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DNA atoms appended to stream cells and clonally enhanced through scaffold intensification are perused out bit by bit adding fluorescent-marked nucleotides. The yield of Illumina sequencing runs is presently the most elevated while being most expense productive simultaneously.
A method that attempts to match this position is Ion Torrent’s semiconductor sequencing. With no optical gadgets, changes in the particle focus because of the arrival of protons from nucleotides which are consolidated into a DNA strand by polymerase are estimated. The read length of every one of these methods is restricted to 200–1000 bps. The third era of sequencing methods disposed of all PCR ventures during library arrangement, subsequently empowering single-particle sequencing.
PacBio’s single-atom constant sequencing records the outflow from phospho-connected nucleotides while fused into DNA strands. Normal read lengths of ~8 kbp are achieved by these sequencers, and epigenetically changed nucleotides can be distinguished at the same time. Superfast nanopore sequencing as acknowledged in Oxford Nanopore Technologies MinION permits the identification of nucleotides while being tightened through a natural pore empowering continuous genome sequencing with simply taking care of, opening up additional opportunities. Indeed, even still mistake inclined, read lengths are in principle just restricted by chromosome length with nanopore sequencing.
Single-cell sequencing to understand the pathogen response
Host-pathogen interactions, particularly in the context of bacterial infections, are dynamic exchanges where transcriptional heterogeneity from both the host and the pathogen can lead to many diverse outcomes through distinct molecular pathways. Transcriptional profiling at the single-cell level, on a genome-wide scale, has enabled a greater appreciation of the cellular diversity in complex biological organisms and the myriad of host transcriptional states during infection.
Here, we highlight recent reports of single-cell RNA sequencing within the context of host-pathogen interactions, describe current limitations for detecting and profiling the transcriptome of invading pathogens at the single-cell level, and suggest exciting future prospects for this technology in the study of infection. We propose that understanding infection as an integrated process between pathogen and host with a resolution at the single-cell level will ultimately inform the development of vaccines with greater productive and protective host immunity, enable the development of novel therapeutics that harness host mechanisms, and yield more accurate biomarkers to guide better diagnostics. (Penaranda, 2019).