Since all the nucleotides are incorporated one at a time, homopolymer estimation is not required, which garners an advantage in sequencing accuracy 8, 9. Both of the attributes are critical to clinical sequencing. Illumina’s SBS offers two distinct advantages over the other SBS platforms: high accuracy and an integrated and automated workflow. Second, the accuracy of homopolymer-calling needs to be improved.
First, DNA amplification using emulsion PCR in oil droplets is cumbersome and difficult to automate and integrate.
#MY BSU SSB TORRENT#
However, to be widely used for targeted sequencing in the clinical space, there are two outstanding concerns with Ion Torrent technology. Using semiconductor sensors for detection can make the platform highly scalable and cost effective. The usage of natural nucleotides results in several advantages including lower cost and longer reads. Ion Torrent’s SBS method detects the byproduct of natural nucleotide incorporation, H +, by ISFET sensors 7.
The most commonly used targeted sequencing methods are based on Sequencing by Synthesis (SBS), including those from Ion Torrent and Illumina. Therefore, targeted sequencing that allows researchers and clinicians to focus on specific gene panels is most widely used because of its low cost and easier data interpretation. Whole-genome sequencing is still relatively expensive, slow and the data are too complex to interpret for routine clinical applications 6. For clinical sequencing, the most important metrics are cost, accuracy, fast turnaround time (TAT) and integrated sample to answer workflow. One of the most important emerging applications of NGS is clinical sequencing, which promises to have a profound impact to human health. Multiple NGS technologies and platforms coexist and each offers unique advantages and disadvantages in sequencing metrics including speed, accuracy, read length, scalability, costs of instrument and reagents and simplicity of workflow 2, 5. Next-generation sequencing (NGS) technologies have drastically reduced the cost of sequencing and placed a wide range of genomic analyses within the capabilities of many laboratories 1, 2, 3, 4. We discuss the advantages of this sequencing chemistry for targeted sequencing as well as its limitations for other applications. This ensemble SBS chemistry has been implemented on an existing Illumina sequencing platform with integrated cluster amplification. This sequencing chemistry uses natural nucleotides and is based on real-time recording of the differential polymerase/DNA-binding kinetics in the presence of correct or mismatch nucleotides. Here we demonstrate a version of the Sequencing by Synthesis (SBS) chemistry that potentially can become a preferred targeted sequencing method in the clinical space. Routine clinical use of targeted NGS mandates inexpensive instruments, fast turnaround time and an integrated and robust workflow. In the clinical space, targeted sequencing has the advantage of allowing researchers to focus on specific genes of interest. However, whole-genome sequencing is still costly and complex for diagnostics purposes. Next-generation sequencing (NGS) has transformed genomic research by decreasing the cost of sequencing.