Next generation sequencing (Illumina Genome Analyzer technique (Complete…
Next generation sequencing
Illumina Genome Analyzer technique
Align data - The data are aligned and compared to a reference, and sequencing differences are identified
Sequencing over multiple chemistry cycles - The sequencing cycles are repeated to determine the sequence of bases in a fragment, one base at a time
Image second chemistry cycle - After laser excitation, the image is captured as before, and the identity of the second base is recorded
Determine second base -The next cycle repeats the incorporation of four labeled reversible terminators, primers, and DNA polymerase
Image first base - After laser excitation, the emitted fluorescence from each cluster is captured and the first base is identified
Determine the first base -The first sequencing cycle begins by adding four labeled reversible terminators, primers, and DNA polymerase.
Complete Amplification - Several million dense clusters of double-stranded DNA are generated in each channel of the flow cell.
Prepare Genomic DNA Sample - randomly fragment genomic DNA and ligate adapters to both ends of the fragments
Denature the double-stranded molecules - Denaturation leaves single-stranded templates attached to the substrate
Attach DNA to surface - bind single-stranded fragments randomly to the inside surface of the flow cell channels
Fragments become double-stranded -The enzyme incorporates nucleotides to build double-stranded bridges on the solid-phase substrate.
Bridge Amplification - Add unlabeled nucleotides and enzyme to initiate solid-phase bridge amplification
Some of the machine has two lanes cannot apply different samples and only one lane mix can be applied in the entire flowcel ("illumina (NovaSeq / HiSeq / NextSeq)" 2018)
Some of the machine use two dyes. This does not give any practical differences in terms of the data quality ("illumina (NovaSeq / HiSeq / NextSeq)" 2018)
Its workflow enables even small lab to have sequencing capabilities ("ILumina Genome Analyzer", 2010).
It uses powerful software that can quickly process from image capture to interactive analysis.
It is flexible, raw and accurate.
It gives the power to comprehensively characterize the genome with both short-insert-paired-end and long-insert-paired-end libraries ("ILumina Genome Analyzer", 2010).
It is accessible to most laboratory
It gives faster results with proven scalability.
Illumina Genome Analyzer is easy to use and produce data efficiently by reducing the time
Illumina Genome Analyzer can produce vast number of precise sequencing reads
Highly parallel, many sequencing reactions take place at the same time.