Methods and systems for using built-in standard curve to measure molecular numbers of biological components

摘要

Methods and devices are disclosed, which perform absolute quantification assay of functional components of biological specimens. The present application relates to manufacture built-in standard curve(s) and computational analysis. The present application further relates to methods for the measurements of such biological components, for example, biochips, PCR arrays, microarrays, and ELISA array.

主权项

1. A method of determining the absolute quantity of a molecule of interest (sample target), comprising:
i. providing a first molecule as control partner 1 and a second molecule as sample probe; ii. providing a third molecule as control partner 2 and a fourth molecule as sample target; iii. providing a series of control partner 1 with different and known copy numbers; iv. attaching the series of control partner 1 onto a solid support at separate locations; v. attaching at least one sample probe or at least one sample target onto the solid support; vi. contacting at least one control partner 2 with the series of control partner 1, wherein the series of control partner 1 recognize and bind the at least one control partner 2, resulting directly or indirectly in a series of detectable control signals, wherein the intensity of each location of the detectable control signals correlates with the known copy number of the control partner 1 in the series of control partner 1, wherein the copy number of control partner 2 is no less than the copy number of control partner 1; vii. detecting the control signals and making a standard curve relating the signal intensities of the series of detectable control signals with the known copy number of the control partner 1 in the series of control partner 1; viii. adding at least one sample target or at least one sample probe to allow the at least one sample probe contacting with the at least one sample target, wherein the at least one sample probe recognizes and binds the at least one sample target, resulting directly or indirectly in a detectable sample signal, wherein the intensity of each location of the detectable sample signals correlates with the copy number of the at least one sample target, wherein the copy number of sample probes is no less than the copy number of sample targets; ix. determining the copy number of the at least one sample target by using the standard curve to translate the sample signal intensity to the copy number of the at least one sample target.