The present invention relates to the technical field of biological detection, and particularly to a PCR base and a PCR instrument.
A polymerase chain reaction (PCR) is a molecular biological technology used for magnifying and amplifying a specific DNA fragment. In the PCR, DNA denatures into a single chain at a high temperature of 95° C. in vitro, a primer is combined with the single chain according to a complementary base pairing rule at a low temperature (usually around 60° C.), and then, DNA polymerase synthesizes a complementary chain along a direction of phosphoric acid to pentose by adjusting the temperature to an optimal temperature (around 72° C.) for the DNA polymerase reaction. An implementation of the PCR generally requires a PCR instrument to control the temperature of each reaction stage and heating time.
To prevent test solution from volatilizing upward in the high-temperature PCR, a hot cover unit is generally covered above a well plate of the test solution by the PCR instrument to reduce volatilization of the test solution with the high temperature above the well plate. The PCR well plate which is usually made of plastic will expand greatly after being heated, but a PCR base that is generally made of metal and used for placing the PCR well plate has an extremely small thermal expansion rate. As shown in
To overcome the above shortcomings of the prior art, an object of the present invention is to provide a PCR base and a PCR instrument.
To achieve the above object and solve the technical problem of the present invention, a technical solution adopted by the present invention is described as follows: a PCR base includes a metal block, wherein the metal block is provided with a plurality of well grooves for holding tubes on a PCR well plate; under an ambient condition of a room temperature being 22±2° C., a well spacing of the PCR well plate is d0, a well spacing of the PCR base is d1, the well spacing d1 of the PCR base is greater than the well spacing d0 of the PCR well plate, and d1=d0+Δd, where Δd refers to a thermal expansion compensation value.
Compared with the prior art, in the present invention, the well spacing d0 of the heated PCR well plate extends to be close to the well spacing d1 of the PCR base during the PCR. The well grooves of the PCR base no longer limit expansion and extension of the PCR well plate, a surface of the PCR well plate is in a planar state, and an upper hot cover can be well covered on the surface of the PCR well plate. Therefore, evaporation and cross-contamination of test solution in the PCR well plate are prevented, heating uniformity of each well position of the PCR well plate is improved, and accuracy of PCR detection is increased.
Further, the thermal expansion compensation value is Δd=ΔT*s*d0, where ΔT refers to a temperature difference value being 20-80 K, and s refers to a thermal expansion coefficient of a material of the PCR well plate and is (30−250)*10−6 mm/K.
Further, the well spacing d1 of the PCR base is (1+0.06%)d0≤d1≤(1+2%)d0.
By adopting the above preferable solution, the thermal expansion compensation value with wide applicability is obtained according to a common material expansion rate of the PCR well plate and a large number of pilot experiments, thereby effectively eliminating arching deformation of the heated PCR well plate and considering universality of the PCR base at the same time.
A PCR instrument includes:
the above PCR base, on which a PCR well plate is placed;
an upper heating unit, including a hot cover and a heating element, wherein the heating element is used to heat the hot cover, and the hot cover is covered on an upper surface of the PCR well plate during a PCR; and
a lower temperature controlling unit, disposed below the PCR base and used to perform temperature control for the PCR base.
By adopting the above preferable solution, the heated PCR well plate no longer deforms, and the hot cover can be matched with the surface of the PCR well plate better.
Further, a first driving mechanism for driving the upper heating unit to ascend and descend is included.
Further, a supporting block is disposed on the PCR base corresponding to an edge below a lip side of the PCR well plate, a vertical guide post is disposed at the bottom of the supporting block, a guide hole matched with the vertical guide post is disposed on the PCR base, and a second driving mechanism for driving the supporting block to move up and down is included.
Further, the second driving mechanism includes a beveled block, a horizontally disposed lead screw pair and a drive motor, the supporting block is mounted on a bevel of the beveled block, the beveled block is connected with a nut of the lead screw pair, and the drive motor drives a lead screw of the lead screw pair to rotate.
By adopting the above preferable solution, the second driving mechanism pushes against the supporting block before the PCR well plate is placed, the PCR well plate is supported on the supporting block by means of its lip side at the edge, and the PCR well plate is not completely placed in the well grooves of the PCR base, thereby effectively preventing concave deformation of the PCR well plate mounted on the PCR base before thermal expansion. After the PCR well plate is heated up to expand, the second driving mechanism then drives the supporting block to move down, tubes of the PCR well plate smoothly enter the well grooves of the PCR base, and then, the first driving mechanism drives the hot cover to cover on the upper surface of the PCR well plate for performing the PCR. After the PCR is ended, the first driving mechanism drives the hot cover to move up, and the second driving mechanism then drives the supporting block to move up to push against the PCR well plate so as to facilitate taking out the PCR well plate and also prevent the tubes from being clamped in the well grooves of the PCR base after the PCR well plate is cooled to contract.
In order to describe technical solutions in embodiments of the present invention more clearly, drawings to be used in the embodiments or descriptions of the prior art will be briefly introduced below. Obviously, the drawings in the following descriptions are merely some embodiments of the present invention, and those of ordinary skill in the art can also obtain other drawings according to these drawings without paying creative work.
Names of corresponding components indicated by numerals and letters in the drawings are described as follows:
1—PCR well plate; 2—PCR base; 3—upper heating unit; 31—hot cover; 32—heating element; 4—lower temperature controlling unit; 41—cooling sheet; 42—water cooling plate; 5—supporting block; 51—vertical guide post; 6—second driving mechanism; 61—beveled block; 62—lead screw pair; 63—drive motor; 91—PCR base; 92—PCR well plate; 93—hot cover.
Technical solutions of embodiments of the present invention will be described clearly and fully below in combination with drawings in the embodiments of the present invention. Obviously, the described embodiments are merely part of embodiments of the present invention rather than all embodiments. Other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present invention without paying creative work shall all fall into the protection scope of the present invention.
As shown in
The above technical solution has the following beneficial effects: during the PCR, the well spacing d0 of the heated PCR well plate extends to be close to the well spacing d1 of the PCR base, the well grooves of the PCR base no longer limit expansion and extension of the PCR well plate, a surface of the PCR well plate is in a planar state, and an upper hot cover can be well covered on the surface of the PCR well plate. Therefore, evaporation and cross-contamination of test solution in the PCR well plate are prevented, heating uniformity of each well position of the PCR well plate is improved, and accuracy of PCR detection is increased.
In other embodiments of the present invention, the thermal expansion compensation value is Δd=ΔT*s*d0, where ΔT refers to a temperature difference value being 20-80 K, and s refers to a thermal expansion coefficient of a material of the PCR well plate and is (30−250)*10−6 mm/K.
In other embodiments of the present invention, the well spacing d1 of the PCR base 2 is (1+0.06%)d0≤d1≤(1+2%)d0. The thermal expansion compensation value with wide applicability is obtained according to a common material expansion rate of the PCR well plate and a large number of pilot experiments, thereby effectively eliminating arching deformation of the heated PCR well plate and considering universality of the PCR base at the same time.
A 96-well PCR well plate is taken as an example. When the PCR well plate is made of PP, its thermal expansion coefficient s is 90*10−6 mm/K, and the temperature difference value ΔT is 80 K. Under the ambient condition of the room temperature being 22±2° C., the well spacing of the PCR well plate is d0=9 mm, and the well spacing of the PCR base is d1=(1+80*90*10−6)*9=9.0648 mm. According to the common material and the heating temperature, the well spacing d1 of the PCR base applicable to the 96-well plate is 9.03-9.22 mm.
A 384-well PCR well plate is taken as an example. When the PCR well plate is made of glass fiber ABS, its thermal expansion coefficient is 30*10−6 mm/K, and the temperature difference value ΔT is 20 K. Under the ambient condition of the room temperature being 22±2° C., the well spacing of the PCR well plate is d0=4.5 mm, and the well spacing of the PCR base is d1=(1+30*20*10−6)*4.5=4.5027 mm. According to the common material and the heating temperature, the well spacing d1 of the PCR base applicable to the 384-well plate is 4.5027-4.55 mm.
As shown in
a PCR base 2, on which a PCR well plate (not shown in
an upper heating unit 3, including a hot cover 31 and a heating element 32, wherein the heating element 32 is used to heat the hot cover 31, and the hot cover 31 is covered on an upper surface of the PCR well plate during a PCR; and
a lower temperature controlling unit 4, disposed below the PCR base 2 and used to perform temperature control for the PCR base 2.
The above technical solution has the following beneficial effects: the heated PCR well plate no longer deforms, and the hot cover can be matched with the surface of the PCR well plate better.
The lower temperature controlling unit 4 may perform temperature control by wind cooling or water cooling. As shown in
As shown in
The above embodiments are merely used to describe technical conceptions and features of the present invention so as to enable those of ordinary skill in the art to understand and implement contents of the present invention, but cannot be used to limit the protection scope of the present invention. Equivalent changes or modifications made based on the spirit of the present invention shall all fall in the protection scope of the present invention.
Number | Date | Country | Kind |
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201910087842.8 | Jan 2019 | CN | national |
Filing Document | Filing Date | Country | Kind |
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PCT/CN2019/100901 | 8/16/2019 | WO | 00 |