CN202390442U - Heat transfer detection device based on DNA amplification - Google Patents

Abstract

The utility model relates to a heat transfer detection device based on DNA amplification, including DNA reaction chamber, optic fibre data processing module, thermal control processing module and host processor, the DNA reaction chamber is an airtight space be equipped with the DNA amplification module in the DNA reaction chamber the upper portion of DNA amplification module is provided with the reaction hole, is equipped with prague sensing probe in the reaction hole, prague sensing probe is connected with the optic fibre core, and demodulation module is connected to the optic fibre core, demodulation module with optic fibre data processing module is connected, a comparator is connected to optic fibre data processing module passing signal line, the comparator pass signal line with the host processor is connected. The utility model discloses a fiber Bragg grating and platinum resistance sensor detect respectively, and centralized processing constructs the data control model, and the whole amplification process of simulation DNA reveals the influence factor to the DNA sample, provides the foundation for revealing the objective law of DNA internal reaction.

Description

A kind of heat passage proofing unit based on DNA cloning

Technical field

The utility model belongs to technical field of biological, is specifically related to a kind of heat passage proofing unit based on DNA cloning.

Background technology

A basic Recurrence Process of DNA gene amplification by sex change--annealing--extend three step of reaction and constitute, live through tens basic Recurrence Process after, just can make gene to be amplified amplify 1,000,000 times.In stage, generally at 93 ℃-95 ℃, the double-stranded DNA template forms single stranded DNA to the temperature in DNA sex change stage behind the hydrogen bond rupture under the heat effect in this stage three primitive reactions; The temperature of annealing stage reduces in this stage system temperature generally at 42 ℃-55 ℃, and primer combines with dna profiling, forms local double-stranded; The temperature in extension stage is generally at 70 ℃-75 ℃, at this stage synthetic and dna profiling complementary DNA chain under the enzyme effect.

The step of archaeal dna polymerase being introduced PCR is: the DNA reaction solution is placed in the test tube, and the test tube that reaction solution is housed is placed in each hole of PCR module of reaction chamber, and the gene amplification process is accomplished by sequence of control automatically.Up to the present; Chinese scholars mainly utilizes automatic control technology that the temperature variation of PCR module is controlled on sequence of control; The immanent structure expression of gene that Stephanie J.Culler seminar has utilized the PCR device analysis, but most achievement in research concentrates on some index of improving round pcr and raising PCR equipment.E.T.Lagally adopts the method that improves temperature rate that the DNA cloning process is studied as far back as calendar year 2001; Grover J. and Juncosa R.D studied the elevation rate of conventional PCR equipment in 2008, and employing Digital Control modes such as T.M.H.Lee have improved the reaction control accuracy of PCR.In the clinical application of PCR; A ubiquitous problem is: the state that is arranged in the DNA reaction solution of each hole test tube of module changes with the difference of step of reaction, in this process, and the constantly fracture or generate of the chemical bond of DNA; The suction that is accompanied by, heat release state are also different; How to analyze these change of state rules, to improving the DNA cloning quality and the temperature variation that is placed on test-tube reaction liquid in each hole of module accurately having been controlled direct influence, the prior-art devices complicated operation; Can not accurately detect as required neatly, being applied in has certain limitation in the actual engineering.

Summary of the invention

The utility model overcomes the deficiency of prior art; A kind of heat passage proofing unit based on DNA cloning has been proposed; Said device utilizes fiber Bragg grating sensor and distribution character model thereof; Utilize this characteristic that the data rule of DNA reaction solution is gathered and analyzed, the heat passage rule of analyzing DNA amplification.

The technical scheme of the utility model is: a kind of heat passage proofing unit based on DNA cloning, comprise DNA reaction chamber, fiber data processing module, thermal control processing module, heat-dissipating space and primary processor, and said DNA reaction chamber is an enclosed space; Said heat-dissipating space is positioned at the bottom of said DNA reaction chamber; And isolate with the DNA reaction chamber, in said DNA reaction chamber, be provided with the DNA cloning module, be provided with reacting hole on the top of said DNA cloning module; Be provided with Prague sensing probe in the reacting hole; Said Prague sensing probe is connected with fiber cores, and fiber cores connects a demodulation module, and said demodulation module is connected with said fiber data processing module; Said fiber data processing module connects a comparer through SW; Be provided with sensing unit and heating unit in the bottom of said DNA cloning module, in said heating unit, be provided with the semi-conductor heating chip, below said semi-conductor heating chip, be provided with a tropical resistance zone; In said sensing unit, be provided with platinum sensor; Said platinum sensor is connected with an analog through SW, and said analog is positioned at heat-dissipating space, and analog is connected with said thermal control processing module through a hardware interface unit; Said thermal control processing module is connected with said comparer through SW, and said comparer is connected with said primary processor through SW.

Said semi-conductor heating chip is connected with a heat controller through wire, and said heat controller is positioned at heat-dissipating space, and heat controller is connected with said thermal control processing module through said hardware interface unit.

In heat-dissipating space, be provided with heat abstractor.

The utlity model has following beneficial effect

1) the utility model adopts Fiber Bragg Grating FBG and platinum sensor to detect respectively; Focus on, make up data control model, the whole amplification procedure of analog D NA; Announcement is to the influence factor of dna sample, for the objective law that discloses the DNA internal-response provides foundation.

2) the utility model is set up the temperature variant distribution character of Fiber Bragg Grating FBG, discloses the suction law of heat release in DNA cloning stage, discloses the objective law of DNA internal-response, provides a whole set of Fiber Bragg Grating FBG temperature variant theory and technology basic data.

3) the utility model has been set up the practicable data acquisition modes of DNA cloning process.

4) the utility model has disclosed the reaction rule of DNA, will help the PCR development of equipments, and market outlook and social benefit are huge.

Description of drawings

Accompanying drawing is the skeleton construction synoptic diagram of the utility model.

Among the figure, 1, reacting hole; 2, Prague sensing probe; 3, fiber cores; 4, DNA reaction chamber; 5, demodulation module; 6, fiber data processing module; 7, comparer; 8, primary processor; 9, thermal control processing module; 10, hardware interface unit; 11, heat controller; 12, semi-conductor heating chip; 13, platinum sensor; 14, heat-dissipating space; 15, analog.

Embodiment

The utility model comprises DNA reaction chamber 4, fiber data processing module 6, thermal control processing module 9, heat-dissipating space 14 and primary processor 8, and said DNA reaction chamber 4 is an enclosed space, and said heat-dissipating space 14 is positioned at the bottom of said DNA reaction chamber 4; And isolate with DNA reaction chamber 4, in said DNA reaction chamber 4, be provided with the DNA cloning module, be provided with several reacting holes 1 on the top of said DNA cloning module; Be generally 96 holes; The DNA reaction solution is housed in the reacting hole 1, in some reacting hole 1, is provided with Prague sensing probe 2, the quantity of Prague sensing probe 2 is confirmed according to concrete detection scheme; In the reacting hole that is provided with Prague sensing probe 21, the DNA reaction solution can be housed; Said Prague sensing probe 2 is connected with fiber cores 3, and fiber cores 3 connects a demodulation module 5, and said demodulation module 5 is connected with said fiber data processing module 6; Said fiber data processing module 6 connects a comparer 7 through SW; Be provided with sensing unit and heating unit in the bottom of said DNA cloning module, in said heating unit, be provided with semi-conductor heating chip 12, below said semi-conductor heating chip 12, be provided with a tropical resistance zone; In said sensing unit, be provided with platinum sensor 13; Said platinum sensor 13 is connected with a weighted-voltage D/A converter 15 through SW, and said analog 15 is positioned at heat-dissipating space 14, and analog 15 is connected with said thermal control processing module 9 through a hardware interface unit 10; Said thermal control processing module 9 is connected with said comparer 7 through SW, and said comparer 7 is connected with said primary processor 8 through SW.Said semi-conductor heating chip 12 is connected with a heat controller 11 through wire, and said heat controller 11 is positioned at heat-dissipating space 14, and heat controller 11 is connected with said thermal control processing module 9 through said hardware interface unit 10.In heat-dissipating space 14, be provided with heat abstractor.

When utilizing said fiber Bragg grating sensor detected temperatures, according to coupled mode theory, the foveal reflex wavelength of Fiber Bragg Grating FBG can be expressed as

m _B＞2n _effM (1)

N in the formula _EffBe the effective refractive index of guided mode, M is the cycle of grating.Can find out that by (1) formula the foveal reflex wavelength is relevant with the grating cycle with effective refractive index;

When grating received variation of temperature and influences, its effective refractive index and grating cycle can change thereupon, thereby reflection wavelength also can change, and relational expression is:

Em _B＞2En _effM，2n _effEM

Obtain following formula substitution formula (1)

$\frac{{\mathrm{Em}}_B}{m_B}>\frac{{\mathrm{En}}_{\mathrm{eff}}}{n_{\mathrm{eff}}},$ $\frac{\mathrm{EM}}{M}---\left(2\right)$

The grating cycle varying type and the thermo-optical coeffecient that are caused by thermal expansion effects cause that the effective refractive index varying type is respectively:

Wherein b and y are respectively the thermal expansivity and the thermo-optical coeffecient of optical fiber.

Can get above-mentioned two formula substitution formula (2):

Make b _T＞k _T∑ m _B, then following formula can be written as:

The foveal reflex wavelength m of common optical fiber _B=1200nm, the temperature sensitivity coefficient k _T=7.5x10 ^-6/ C, therefore, b _TIt is the function of temperature.Formula (3) is a linear relationship to certain temperature, but when temperature variation was big, above-mentioned linear formula showed nonlinear characteristic.Therefore be adapted at the bigger polymeric materials of outer wrap temperature sensitivity coefficient, as, k _T=87x 10 ^-6/ C, at this moment, b _T=9.88, that is, for the spatial resolution of every degree between 0-9.88.

The utility model adopts the conventional modular unit in 96 holes; DCU data control unit divides three tunnel placement sensor groups, and every road is provided with four platinum sensors 13, and the control data collecting unit is used to make an experiment; Can be under the situation of condition permission; In the reacting hole space on module top, arrange the platinum sensor 13 in 12 reaction chambers, be called for short the reaction chamber transmitter, be used for the homogeneous and the harmony of acquisition module temperature with the correspondence position of platinum sensor 13; The data acquisition unit of setting up is used for experimental system is carried out data analysis, and as the auxiliary basic data of Bragg grating DAS.

Suppose that platinum sensor 13 is A _Ij(i=1-3, j=1-4), the reaction chamber transmitter is W _Ij(i=1-3, j=1-4),

Wherein, A _Ij(i=1-3, j=1,2) and W _Ij(i=1-3 j=1-4) is analog quantity, and establishing the function that analog quantity transfers data volume (decimal system temperature value) to is G, can get through after experiment and the The Coupling: t at any one time, and the temperature data of its platinum sensor acquisition is:

$\begin{array}{c}{T}_{\mathrm{ij}}\left(t\right)=\\ W_{\mathrm{ij}}\left(t\right)[1,\stackrel{3}{\underset{i>1}{\mathrm{\Σ}}}(\underset{j>1}{\overset{4}{\mathrm{\Σ}}}(1,\sqrt{1,{({W^2}_{\mathrm{ij}}\left(t\right).{A^2}_{\mathrm{ij}}\left(t\right))}^2})/\sqrt{1,({W^2}_{\mathrm{ij}}\left(t\right),{A^2}_{\mathrm{ij}}\left(t\right))})W_{\mathrm{ij}}\left(t\right))/4]\end{array}---\left(4\right)$

Wherein, i=1-3, what j=1-4, following formula represented is the actual temperature value of certain TP of automatic control system loading.

T at any one time, the m of optical fiber _BValue is constantly to change, the data of the DNA reaction solution of the dna sample that foundation is measured, and foundation (3) Shi Kede:

Can know that from (5) formula the temperature variation ET of the DNA reaction solution between t moment partial zones in (Et) can pass through to calculate Em according to the demodulated equipment of Bragg grating _B(t) obtain, and temperature value can be by the three-way sensor combined calculation of module temperature constantly at arbitrary t.

So, t at any one time ₀, the actual temperature data that its automatic control system loads can be calculated by (4) formula, $T_{\mathrm{Ij}}\left(t_0\right)=$

$W_{\mathrm{ij}}\left(t_0\right)[1,\underset{i>1}{\overset{3}{\mathrm{\Σ}}}(\underset{j>1}{\overset{4}{\mathrm{\Σ}}}(1,\sqrt{1,{({W^2}_{\mathrm{ij}}\left(t_0\right).{A^2}_{\mathrm{ij}}\left(t_0\right))}^2})/\sqrt{1,({W^2}_{\mathrm{ij}}\left(t_0\right),{A^2}_{\mathrm{ij}}\left(t_0\right))})W_{\mathrm{ij}}\left(t_0\right))/4]$

Wherein, i=1-3, j=1-4.Can know that from top analysis between the partial zones of t0 till constantly, the variable quantity of the actual temperature of the corresponding TP in its reaction chamber can be calculated by (5) formula:

ET(T ₀)| _ij＞Em _B(t ₀)b _T(b _TΥ0)

Comparison through (4) formula and (5) formula can accurately calculate at arbitrary t ₀Constantly, the precise temp changing value Ei of DNA reaction chamber internal reaction liquid _Ij| t ₀, that is,

Ei _ij|t ₀＞|ET(t ₀)| _ij·T _ij(t ₀)|(6)

Can extrapolate the heat absorption of DNA reaction solution reaction process and the concrete rule of heat release from (6) formula, and can accurately simulate DNA biological respinse characteristic, and then set up the data model of DNA reaction, according to the data T that obtains according to above-mentioned formula _Ij(t), through confirming the DNA step of reaction and inhale the heat release state with the contrast of ET.

Claims (3)

1. heat passage proofing unit based on DNA cloning, it is characterized in that: comprise DNA reaction chamber, fiber data processing module, thermal control processing module, heat-dissipating space and primary processor, said DNA reaction chamber is an enclosed space; Said heat-dissipating space is positioned at the bottom of said DNA reaction chamber; And isolate with the DNA reaction chamber, in said DNA reaction chamber, be provided with the DNA cloning module, be provided with reacting hole on the top of said DNA cloning module; Be provided with Prague sensing probe in the reacting hole; Said Prague sensing probe is connected with fiber cores, and fiber cores connects a demodulation module, and said demodulation module is connected with said fiber data processing module; Said fiber data processing module connects a comparer through SW; Be provided with sensing unit and heating unit in the bottom of said DNA cloning module, in said heating unit, be provided with the semi-conductor heating chip, below said semi-conductor heating chip, be provided with a tropical resistance zone; In said sensing unit, be provided with platinum sensor; Said platinum sensor is connected with an analog through SW, and said analog is positioned at heat-dissipating space, and analog is connected with said thermal control processing module through a hardware interface unit; Said thermal control processing module is connected with said comparer through SW, and said comparer is connected with said primary processor through SW.

2. the heat passage proofing unit based on DNA cloning according to claim 1; It is characterized in that: said semi-conductor heating chip is connected with a heat controller through wire; Said heat controller is positioned at heat-dissipating space, and heat controller is connected with said thermal control processing module through said hardware interface unit.

3. the heat passage proofing unit based on DNA cloning according to claim 1 and 2 is characterized in that: in heat-dissipating space, be provided with heat abstractor.

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