CN2306230Y

CN2306230Y - Semiconductor gene magnification chamber

Info

Publication number: CN2306230Y
Application number: CN 97213998
Authority: CN
Inventors: 王志西
Original assignee: Individual
Current assignee: Individual
Priority date: 1997-04-18
Filing date: 1997-04-18
Publication date: 1999-02-03
Anticipated expiration: 2007-04-18

Abstract

The utility model relates to a gene amplification chamber used for a PCR technology. The utility model is composed of a semiconductor thermopile, a heat conductive working board, a radiation board, etc., and the semiconductor thermopile of the utility model is formed by the overlapping of multiple layers of semiconductor thermopiles. At any time, the contact surfaces of two adjacent layers of semiconductor temperature difference blocks are in opposite states of cold and hot. The utility model brings possibility that the ordinary semiconductor difference temperature blocks can be applies to a PCR gene augmentor. The utility model has the advantages of high precision in temperature control, long service life, low cost and fast speed in raising and reducing temperature.

Description

A kind of semiconductor-based gene-amplification chamber

The utility model relates to a kind of improved gene-amplificative instrament parts, belongs to biological experiment technical equipment, particularly molecular biology experiment test set field.

Polymerase chain reaction (Polymerase chaim Reaction) is called for short round pcr, is a kind of new technology of simulating the n DNA reproduction process at external rapid amplifying dna segment.Round pcr is easy, quick, sensitive with it, the specificity advantages of higher, is developed rapidly in fields such as life science, medical diagnosis, genetic engineering, legal medical expert and archeology and uses.

The round pcr principle is template DNA, four kinds of deoxymononucleotides (DNTP) solution, thermotolerance polymerase, the Mg that adds an amount of damping fluid, trace in Eppendorf tube ²⁺Primer with two synthetic DNAs.At first with above-mentioned solution heating, the template DNA two strands is dissociated, form single stranded DNA, this is called the sex change of DNA, removes after the condition of sex change, and the strand of sex change can recombine get up, and forms two strands, and this is called the DNA renaturation, also cries annealing.Template DNA is decomposed into two strands, then through after high-temperature denatured in the pcr amplification system, system temperature reduces, and primer annealing combines with mutual complementary template DNA, form partial two strands, this is the starting point of dna replication dna, i.e. fixedly starting point of Yan Shening, temperature of reaction with solution rises to middle temperature again, under the effect of polymerase, is raw material with DNTP, with primer is the starting point of duplicating, a two strands of template DNA forms two two strandss after unwinding and annealing, this is called extension.So repeat to change temperature of reaction, promptly high-temperature denatured, low-temperature annealing, extend three phases with middle temperature, become a circulation again, through a circulation gene copy number of special section is put and be twice, general sample finally makes the gene number amplify millions of times through 30 circulations.

High-temperature denatured, low-temperature annealing, middle temperature are extended in the three phases, and temperature requirement is: high-temperature denatured 90 ℃～97 ℃, anneal about 50 ℃, and extend about 70 ℃.When temperature variation, require the lifting gradient of temperature very big, it is steep that the curvilinear motion of temperature is wanted, and requires temperature control very accurate simultaneously, for a gene-amplificative instrament, how to realize that the precision circulation of temperature just becomes the core of its technology.

The pcr gene amplification instrument that occurs on market at present has following several types by its heating-cooling principle:

1. with the abundant pot of the water of three differing tempss, movement of sample is reached the PCR temperature cycle by mechanical manipulator.The mechanical manipulator of this type damages easily, and temperature variation is few, can not adapt to the requirement that various temperature changes.

2. reach the PCR temperature cycle with recycle gas or liquid, this instrument temperature control is poor, and speed of response is slow.

With resistance element heating, in conjunction with the pcr amplification instrument of compressor cooling, this type of model-performance is stable, temperature control is accurate, but speed of response is slower, and costs an arm and a leg.

4. the semiconductor material of selecting for use according to bohr subsides effect comes heating-cooling to finish pcr amplification.Advantages such as because it is fast that conductor refrigeration pyrogenicity piece (being temperature difference piece) has cold and hot switching speed, thermosteresis is little, and device is simple, and is in light weight, late nineteen eighties, a lot of in the world companies finish the pcr amplification instrument that the semiconductor temperature difference piece is used in development mutually.

Because a large amount of employing of semiconductor temperature difference piece three telluriumizations two bismuth materials are made, the N type made from this material or the semiconductor material intensity of P type are all very low, its solder flux is non-refractory again, the general unloaded temperature difference only has 60 ℃～66 ℃, and the working condition of pcr amplification instrument requires its load temperature difference to surpass more than 70 ℃, require cold and hot velocity of variation height simultaneously and the cycle little, therefore the temperature difference piece made from common three dysprosiumizations, two bismuth semiconductor materials is used for pcr amplification instrument life-span weak point, and this has become the unsolved difficult problem in our times various countries.

In recent years, the external up-to-date pcr amplification instrument of producing, temperature rate can reach 3 ℃/second, control is accurate, temperature overshot is no more than 0.1 ℃, be present state-of-the-art pcr gene amplification instrument, but the semiconductor temperature difference piece that this instrument uses is the exotic materials manufacturing, cost is high, and a general import machine price is at 10～240,000 Renminbi.

The purpose of this utility model is to provide a kind of gene amplification chamber that is used for pcr gene amplification instrument of adopting general semiconductor temperature difference piece to make.

The purpose of this utility model is achieved in that

A kind of semiconductor-based gene-amplification chamber, the indoor placement of increasing holds the centrifuge tube 4 that is amplified gene, the heating panel 5 that the gene amplification chamber is piled and joined by semiconductor temperature difference, cooling fan is formed, thermally insulating housing 6 is piled semiconductor temperature difference, heat conduction working plate 3 surrounds within it, its basic improvements are, described semiconductor temperature difference heap is stacked together by two or more semiconductor temperature difference pieces 2, adjacent two layers semiconductor temperature difference piece working face area is identical, performance perameter is also identical, and corresponding one by one overlapping placement, at any one time, the contact surface of adjacent two layers semiconductor temperature difference piece is in cold and hot inverse state.

Above-mentioned semiconductor-based gene-amplification chamber scribbles the heat conduction adipose membrane between the described adjacent two layers semiconductor temperature difference piece.

Above-mentioned semiconductor-based gene-amplification chamber, the number of plies of the semiconductor temperature difference piece that described semiconductor temperature difference heap is stacked is a layer 2-4.

Above-mentioned semiconductor-based gene-amplification chamber, described thermally insulating housing is made by rigid foam.

In the utility model, temperature difference piece stacks, the connection of cold and hot polyphone owing to the semiconductor temperature difference heap that is used for temperature control has adopted, make it can finish the requirement of PCR temperature cycle effectively, the temperature difference that simultaneously makes every layer of semiconductor temperature difference piece be born again is reduced in 40 ℃, much smaller than general semiconductor temperature difference piece temperature difference parameters limits value, thereby solved the life problems of general semiconductor temperature difference piece in pcr gene amplification instrument smoothly, make that general semiconductor temperature difference piece is applied to pcr gene amplification instrument becomes possibility.These gene amplification chamber heating and cooling are quick, the accuracy of temperature control height, and also cost is low, and the pcr gene that uses described amplification chamber to make increases the instrument cost less than 2000 yuans, much smaller than external imported product.

Below in conjunction with accompanying drawing the utility model is elaborated:

Fig. 1 is a structural representation of the present utility model;

Fig. 2 is a vertical view of the present utility model.

As can be seen from Figure 1, the structure of amplification described in the utility model chamber is by semiconductor temperature difference piece 2, heat conduction working plate 3, and heating panel 5, thermally insulating housing 6 is formed, and centrifuge tube 4 and thermometer hole 7 are arranged on heat conduction working plate 3.

Key of the present utility model is the connection and the installation of semiconductor temperature difference piece.It is the semiconductor temperature difference piece of TEC1-12708 that the semiconductor temperature difference piece is selected model for use, divide two superimposed placement up and down, during external power supply wire, the polarity of wiring should make up and down, and the contact surface of two-layer semiconductor temperature difference piece is opposite state of temperature, promptly the upper surface when upper strata semiconductor temperature difference piece is the pyrogenicity face, when lower surface is refrigeration face, it is pyrogenicity that lower floor's semi-conductor also should be upper surface, lower surface is a refrigeration face, the contact surface of upper strata semiconductor temperature difference piece and lower floor's semiconductor temperature difference piece is cold just like this, two kinds of different state of temperatures of heat; When pilot circuit was reverse, the lower surface of upper strata semiconductor temperature difference piece was the pyrogenicity face, and the semi-conductive upper surface of lower floor is than huyashi-chuuka (cold chinese-style noodles), and their state of temperature is opposite forever.

The working area of two-layer up and down semiconductor temperature difference piece is identical, that is the performance perameter of upper and lower two-layer semiconductor temperature difference piece is identical, and corresponding one by one overlapping placement can be satisfied the requirement that semiconductor temperature difference is piled rapid transformation temperature state like this, simultaneously unlikely again damage semiconductor temperature difference piece.Because during the work of this kind semiconductor temperature difference piece, refrigerating capacity is about about 1/3 of pyrogenicity amount, therefore after adopting above-mentioned installation method and wiring, the pyrogenicity of the contact surface of two-layer semiconductor temperature difference piece and chilling effect produce neutralization, it is the semiconductor temperature difference piece surface temperature decline of pyrogenicity face that the result makes contact surface, and the semiconductor temperature difference piece surface temperature of the refrigeration face of making rises.But meanwhile, the temperature head that two-layer conductor temperature piece is born does not change, only for every layer of semiconductor temperature difference piece, the temperature difference between the upper and lower surface of itself bearing has just reduced, make it be lower than the temperature difference limit that general semiconductor temperature difference piece can be born, solved the difficult problem that the unloaded temperature difference limit of general semiconductor temperature difference piece is too small, can not satisfy the job requirement of pcr gene amplification instrument more satisfactorily.

After adopting this structure, the operating voltage that the semiconductor temperature difference piece is required, electric current also can be far smaller than calibration value, has prolonged the work-ing life of semiconductor temperature difference piece greatly.

Scribble the heat conduction adipose membrane between the upper and lower two-layer semiconductor temperature difference piece, such as silicone grease, in order to heat transfer.

In addition, can see from Fig. 1 that working plate 3 contacts with the upper surface of semiconductor temperature difference heap 1.During work, working plate 3 absorbs heat or cooling; Heating panel 5 contacts with the lower surface of semiconductor temperature difference heap, during work, and heating panel 5 heat radiations.

Gene-amplificative instrament generally computerizeds control when work.Computer changes according to parameters such as the voltage of the sequence circuit of establishment and semiconductor temperature difference heap, electric current, polarity, and the feedback signal that receives temperature element adjusts, and makes the temperature of working plate carry out circulation change in strict accordance with the requirement of gene amplification.

The data parameters of a typical gene amplification chamber is:

97 ℃ of denaturation temperatures, 50 ℃ of annealing temperatures, 70 ℃ of elongating temperatures, gradient of temperature speed 〉=3 ℃/second, temperature overshot≤0.1 ℃, the single block of semiconductor temperature difference piece temperature difference: in 40 ℃, single block of semiconductor temperature difference piece voltage: about 8V, be 2/3rds of rated voltage, single block of semiconductor temperature difference piece electric current: about 4A, be 1/2nd of nominal current.

Claims

1. semiconductor-based gene-amplification chamber, the indoor placement of increasing holds the centrifuge tube that is amplified gene, the gene amplification chamber reaches the heating panel [5] that joins with it by the semiconductor temperature difference heap, cooling fan is formed, thermally insulating housing [6] is piled semiconductor temperature difference, heat conduction working plate [3] surrounds within it, it is characterized in that, described semiconductor temperature difference heap [1] is stacked together by two or more semiconductor temperature difference pieces (2), adjacent two layers semiconductor temperature difference piece working area is identical, performance perameter is identical, and corresponding one by one overlapping placement, at any one time, the contact surface of adjacent two layers semiconductor temperature difference piece is in cold and hot inverse state.

2. gene amplification according to claim 1 chamber is characterized in that, scribbles heat-conducting silicone grease between the adjacent two layers semiconductor temperature difference piece (2).

3. gene amplification according to claim 2 chamber is characterized in that, the number of plies of the semiconductor temperature difference piece (2) that described semiconductor temperature difference heap [1] is stacked is a layer 2-4.

4. gene amplification according to claim 3 chamber is characterized in that, described thermally insulating housing (6) is made by rigid foamed.