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Revision as of 12:18, 16 October 2017

Team:NUDT_CHINA

Model

Development of A Novel Blood-MicroRNA Handy Detection System with CRISPR

Abstract

This model is created to evaluate the effectiveness of initial design, and offers guidelines on how the system can (or must) be improved. (You can go to PROJECT. page to see more

Introduction

We create mathematical models of two aspects of our project, a RCA model and a signal detection model.

Assumption and Justification

About model

1. MiRNA is not degraded throughout the reaction process.

2. The two fusion proteins of dCas9 and split-HRP fragments have the same ability to combine with the stem-loop structure, and only when two different proteins next to each other, can they have the ability to catalyze substrate and produce signal.

3. The number of stem-loop structures in each RCA product is equal under a certain reaction time.

4. The enzymatic activity remains unchanged with time under the premise of excessive amount of enzymes or a short-time reaction.

About the data

1. The data we obtain from wet-lab experiment are reliable.

2. All the results are trustworthy in the process of statistical processing and data calculation.

Model

Notations

Symbol

Definition

x

The concentration of miRNA (pM)

C1

The concentration of initiated probe (Abbreviated to iprobe)

k1

A constant representing the scale factor

Km

One of the characteristic constants of phi29 DNA polymerase

Vmax

One of the characteristic constants of phi29 DNA polymerase

k2

A constant representing the scale factor

V

The initial speed of RCA

n1

The moles of RCA product

n2

The number of stem-loop structures in each RCA product

n

The total amount of stem-loop structures

N

The molecule number of the fusion protein of dCas9 and split-HRP fragments

k3

A constant representing the scale factor

y1

The fluorescence intensity of DNA-dye-complex (RFU)

N1

The molecule number of the fusion protein of dCas9 and split-HRP fragments in the solution

N2

The molecule number of the fusion protein of dCas9 and split-HRP fragments binding with stem-loop structure

k4

A constant representing the scale factor

k5

A constant representing the scale factor

ρ

Signal to noise ratio(Abbreviated to SNR)

I

The molecule number of formed intact HRP proteins

I1

The molecule number of formed intact HRP proteins in the solution

I2

The molecule number of formed intact HRP proteins through binding with stem-loop structure

t

Reaction time

y2

The signal intensity (OD450)

Peyto Lake


Figure 1. Schematic diagram