Ystem and method for adaptive reagent control in nucleic acid sequencing
An adaptive, sequencing-reaction technology that is used in molecular biology to address issues such as errors
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[0045] Certain exemplary embodiments of systems and methods involving preparation and processing of samples, generation of sequence data, and analysis of sequence data are generally described below, some or all of which are applicable to embodiments of the invention. In particular, exemplary embodiments of systems and methods and computer systems for preparation of template nucleic acid molecules, amplification of template molecules, generation of target-specific amplicons and / or genomic libraries, sequencing methods and instrumentation are described.
[0046] In typical embodiments, nucleic acid molecules from experimental or diagnostic samples must be prepared and processed from their raw state into template molecules suitable for high-throughput sequencing. Processing methods can vary depending on the application, resulting in template molecules with different properties. For example, in certain embodiments of high-throughput sequencing, it is preferred to generate template...
Embodiment approach
[0077] As noted above, the present invention includes methods for determining the level of activity associated with one or more enzymatic reagents and dynamically adjusting the concentration of one or more of the measured enzymatic reagents to provide significantly optimized levels of reagent activity. In particular, the present invention can be used to reduce introduced errors in sequence data due to delay and incomplete elongation effects due to levels of unwanted nucleotide species in certain process steps, and to adjust available data from the sequencing process. Detection signal output level.
[0078] Certain embodiments of the invention can be used as a tool for calibrating signal output between multiple sequencing instruments and / or different sequencing cycles. Also, signal swings that are too high or too low and the "signal bleed" effect (where the signal from one cycle is so high that the signal persists and bleeds to the next cycle, or in some cases released to adjace...
Embodiment 1
[0110] Example 1 - Validation of the adaptive apyrase process
[0111] Process validation for measuring apyrase activity, determining a pulse width correction factor is required to achieve a predetermined apyrase activity, and adjusting the apyrase pulse by the correction factor.
[0112] Apyrase activity method: the amount of ATP that can be degraded by apyrase can be determined by the enzyme activity in the well;
[0113] Correction factor method: a multiplication factor for pulse width is required to obtain a predetermined apyrase activity;
[0114] Relative signal method: detection of apyrase activity. Correcting the signal generated by pulses of ATP and apyrase concentrations to the signal generated by ATP alone; and
[0115] Setpoint method: desired predetermined relative signal. This is a relative signal and the correction factor should correct for the measured apyrase pulse width.
[0116] Write a script (script) for the process of measuring apyrase activity. The ...
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