Hello,
I am trying to understand the technology of sequencing Oxford Nanopore.
if 1D is sequenced from a single strand. How to build 2D from "1D Temp" and "1D Comp" ??
I speak in the image of step "G" and "H".
The explanation of the entire process is:
(A) Suspended library molecules are concentrated near nanopores embedded in the membrane. A voltage applied across the membrane induces a current through the nanopores. (B) Schematic of a library molecule, showing dsDNA ligated to a leader adapter pre-loaded with a motor protein and a hairpin adapter pre-loaded with a hairpin protein, and the tethering oligos. (C) Sequencing starts from the 5’ end of the leader adapter. The motor protein unwinds the dsDNA allowing single-stranded DNA to pass through the pore. (D) A flow cell contains 512 channels (grey), each channel consisting of 4 wells (white). Each well contains a pore (blue) and a sensor. At any given time, the device is recording the data stream from the wells of the active well-group, in this example, g1. (E) Perturbation in the current across the nanopore is measured 3,000 times per second as ssDNA passes through the nanopore. (F) The ‘bulk data’ are segmented into discrete ‘events’ of similar consecutive measurements. The 5-mer corresponding to each event is inferred using a statistical model. (G) The 1D base-calls are inferred separately for the template and complement event signals. (H) Alignment of the 2D base-calls from the event signals from both, and the 1D base-calls are used to constrain the 2D base-calls.
I am trying to understand the technology of sequencing Oxford Nanopore.
if 1D is sequenced from a single strand. How to build 2D from "1D Temp" and "1D Comp" ??
I speak in the image of step "G" and "H".
The explanation of the entire process is:
(A) Suspended library molecules are concentrated near nanopores embedded in the membrane. A voltage applied across the membrane induces a current through the nanopores. (B) Schematic of a library molecule, showing dsDNA ligated to a leader adapter pre-loaded with a motor protein and a hairpin adapter pre-loaded with a hairpin protein, and the tethering oligos. (C) Sequencing starts from the 5’ end of the leader adapter. The motor protein unwinds the dsDNA allowing single-stranded DNA to pass through the pore. (D) A flow cell contains 512 channels (grey), each channel consisting of 4 wells (white). Each well contains a pore (blue) and a sensor. At any given time, the device is recording the data stream from the wells of the active well-group, in this example, g1. (E) Perturbation in the current across the nanopore is measured 3,000 times per second as ssDNA passes through the nanopore. (F) The ‘bulk data’ are segmented into discrete ‘events’ of similar consecutive measurements. The 5-mer corresponding to each event is inferred using a statistical model. (G) The 1D base-calls are inferred separately for the template and complement event signals. (H) Alignment of the 2D base-calls from the event signals from both, and the 1D base-calls are used to constrain the 2D base-calls.
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