Methods and applications for target quantification

1. A method of quantifying a target in an experimental sample comprising:

a) measuring a nucleic acid detection assay signal at a plurality of time intervals from an experimental sample and at least two standard control samples containing known target copy numbers that are different, thereby generating signal data;

b) providing said signal data to a computer system, wherein said computer system has stored therein a target quantifying software application;

c) processing said signal data with said target quantifying software application such that said target quantifying software:

i) generates a threshold signal line having i) an initial signal value at time zero, and ii) a threshold slope; wherein said threshold signal line is above background level and intersects said experimental curve and two or more of said at least two standard control curves;

ii) processes said signal data and said threshold signal line to generate an experimental intersecting time value and at least two standard control intersecting time values;

iii) processes said at least two standard control intersecting time values and said known target copy numbers to generate log plot data comprising coordinates for a log/log plot, or a linear/log plot, of said intersecting time values versus said known target copy numbers;

iv) processes said log plot data to generate a slope equation describing a resulting slope fit to said log plot data; and

v) processes said experimental intersecting time value with said slope equation to generate a quantified target copy number for said experimental sample; and

d) transmitting said quantified target copy number to a user interface display.

2. The method of claim 1 , wherein said threshold slope of is zero, or wherein said threshold slope is negative.

3. The method of claim 1 , wherein said nucleic acid detection assay comprises an invasive cleavage assay.

4. The method of claim 1 , wherein said nucleic acid detection assay is performed under isothermal conditions.

5. The method of claim 1 , wherein said target comprises a nucleic acid sequence.

6. The method of claim 5 , wherein said nucleic acid sequence comprises a micro-RNA or siRNA sequence.

7. A system for quantifying a target in an experimental sample comprising:

a) a device configured to detect assay signal level at a plurality of time intervals from an experimental sample and at least two standard control samples containing known target copy numbers that are different, to generate initial signal data;

b) a target quantifying software application configured to:

i) process said initial signal data to generate signal data that is capable of being plotted as an experimental curve and at least two standard control curves;

ii) generate a threshold signal line having i) an initial signal value at time zero, and ii) a threshold slope; wherein said threshold signal line is above background, and intersects said experimental curve and two or more of said at least two standard control curves;

iii) process said signal data and said threshold signal line to generate an experimental intersecting time value and at least two standard control intersecting time values;

iv) process said at least two standard control intersecting time values and said known target copy numbers to generate log plot data comprising coordinates for a log/log plot, or linear/log plot, of said intersecting time values versus said known target copy numbers;

v) process said log plot data to generate a slope equation describing a resulting slope fit to said log plot data; and

vi) process said experimental intersecting time value with said slope equation to generate a quantified target copy number for said experimental sample; and

c) a computer system having stored therein said target quantifying software application, wherein said computer system comprises computer memory and a computer processor.

8. The system of claim 7 , wherein said target comprises a nucleic acid sequence.

9. The system of claim 8 , wherein said nucleic acid sequence comprises an RNA or DNA sequence.

10. The system of claim 7 , wherein said assay signal level is generated by a nucleic acid detection assay.

11. The system of claim 10 , wherein said nucleic acid detection assay comprises an invasive cleavage assay.

12. The system of claim 10 , wherein said nucleic acid detection assay is configured to run under isothermal conditions.