DE202006001483U1 - Device for automatic production of agarose gel composition, comprises interchangeable supply containers, reception container exhibiting an outlet and a controllable and/or adjustable arrangement, and an operating unit and/or a display - Google Patents

Abstract

The device for automatic production of agarose gel composition for electrophoretic separation of nucleic acid, comprises a first interchangeable agarose supplying container (2), second interchangeable buffer (21) supplying container (3), a third interchangeable- and temperable supply container (5) for the supply of a color solution, an operating unit (14) and/or a display, and an interchangeable reception container (4) exhibiting an outlet and a controllable- and/or regulatable arrangement to discharge a part of the container contents from the reception container. The device for automatic production of agarose gel composition for electrophoretic separation of nucleic acid, comprises a first interchangeable agarose supplying container (2), second interchangeable buffer (21) supplying container (3), a third interchangeable- and temperable supply container (5) for the supply of a color solution, an operating unit (14) and/or a display, and an interchangeable reception container (4) exhibiting an outlet and a controllable- and/or regulatable arrangement to discharge a part of the container contents from the reception container. The first and the second supply containers show a controlling- and/or regulating unit for supplying a defined quantity of agarose in the reception container and are assigned as a cooling- and/or heating equipment and as an equipment for cooling the reception container. A programmable controlling- and/or regulating unit is connected with the controllable and/or regulatable arrangement. The heating equipment comprises electrical heater, Peltier heater, fuel heater, radiation heater and/or microwave heater. The cooling equipment comprises Peltier cooler, refrigerator, and circulation cooler with compressor, a direct- and/or an indirect cooler. The supply containers are connected with the reception container by a pipeline. A part of the device is isolated. The heating equipment is arranged in such a manner that the sections of the pipelines are heatable. The controllable and/ or adjustable arrangement shows a valve and/or a pump. The supply containers and/or the reception container are assigned as a weighing device, a flow meter, a level meter or an agitating device.

Description

The The invention relates to a device for automated production of agarose gel compositions.

The Production of agarose gels, for example for electrophoretic Separation of nucleic acids takes a lot of time, especially in molecular biological laboratories Claim. That's how it often happens before that Gelrezepturen searched, Agaroseanteile computes, individual Chemicals collected and also individual buffer solutions new have to be recognized before the components can be combined to form an agarose gel. To the joining Finally, the individual gel components consist of the boil-up of the agarose gel compositions still the danger of overcooking the agarose gel composition. This almost everyday Procedures are not only very time consuming but result due to from variances in the individual work steps to not always uniform Agarose gels.

aim The present invention is therefore an apparatus for automated to water of agarose gels which provide a rapid and reproducible Producing agarose gel compositions enabled.

Is solved the object by the features of claim 1. refinements The invention are the subject of the respective subclaims.

According to the invention is an apparatus for the automated production of agarose gel compositions, with a first storage vessel for provision in particular powdered agarose, a second storage vessel for the provision of a Buffer, in particular a TAE or a TBE buffer, and a Receptacle provided, wherein the receptacle a heating device assigned. The first storage vessel and the second storage vessel contain each one tax and / or adjustable device for feeding a defined amount in the receptacle. The receptacle points an outlet and a controllable and / or controllable device on to a part of the vessel contents to be able to discharge from this. Furthermore, a programmable control and / or regulating unit is provided which associated with the tax and / or controllable facilities.

The arranged on the receptacle heater serves to agarose after adding the buffer from the second Storage vessel by increasing the Temperature over 80 ° C in Solution and thus in their liquid State to convict. Thereby that the first storage vessel and the second storage vessel respectively Controllable and / or controllable devices for supplying a defined amount contained in the receptacle, Is it possible, their content is at least partially regulated and / or controlled in to transfer the receptacle. Under a defined amount in the present invention is the amount meant by agarose and buffer, which is required to the desired amount on agarose gel composition with user selected concentrations to produce agarose and buffer. The agarose content of the finished Agarose gel composition is variable and can be in one range from about 0.6% to about 4%.

Thereby, the receptacle comprises an outlet, which a controllable or controllable entity, such as a Valve and / or a pump, is attached, may be part of the vessel contents of the receptacle and / or be made controllable from this. So it is possible to one selectable Volume of the gel composition from the receptacle in a Chamber for casting of agarose gels. Also can in the receptacle a larger volume as that to the filling needed the gel chamber Volume of agarose gel composition can be prepared to from it pour several agarose gels.

The Control and / or regulating unit, which the process steps of the device in particular after User's preferences controls, regulates and monitors, allows automates the desired Adjust agarose concentration in the agarose gel and the agarose gel composition with one adapted to the volume of the gel chamber used volume manufacture.

In Advantageously, the first storage vessel and / or the second storage vessel is a device for cooling and / or heating and / or the receptacle associated with a device for cooling. By the cooling the device can increase the durability of the solutions used, Starting materials and gels over a longer one Significantly increased over time become. For example, it is advantageous to the buffer or the agarose during prolonged Standstill of the device to cool to prevent possible growth of microorganisms such as bacteria and mushrooms. As a result, impurities in the Gelled by unsterilies be minimized and the maintenance intervals of the device according to the invention be enlarged.

by virtue of the heatable configuration of the storage vessels, it is possible to the TAE buffer or the Preheat TBE buffer, whereby a faster production of the agarose gel composition in the Receiving receptacle reached can be.

In the present invention, the Hei For example, it may be an electric fuel heater, a Peltier heater, a radiant heater, and / or a microwave heater, and serves, for example, to agitate the agarose gel composition into its liquid state during mixing in the receptacle by heating to temperatures greater than about 80 ° C and to maintain the agarose gel composition in this liquid state at temperatures of about 45 ° C to about 60 ° C.

at the cooling device it may be, for example, a Peltier cooling, a chiller, a circulation cooling with compressor, a direct cooling and / or indirect cooling act.

According to one another embodiment The invention relates to the first storage vessel for receiving the agarose and the second storage vessel for receiving of the buffer Lines, in particular over Pipelines with the receptacle in which the starting materials are mixed together. The lines, in particular the pipelines are advantageously designed such that Dead zones, especially flow dead zones, in which the agarose gel composition could harden, largely avoided become. In this context, the lines should also be interpreted in such a way that the connections between the individual vessels as short as possible and without corners are designed.

in the Within the scope of the invention, it has proven to be advantageous that the device has a heating device which is designed in this way is that with this at least sections of the lines are heated. For example, the outlet of the receptacle via a Heating coil of the heating of the receptacle designed heated be, thereby curing the agarose gel composition in this pipeline by maintaining a temperature above about 45 ° C in the agarose gel composition can be largely avoided.

By thermal insulation, at least a portion of the device, in particular the reservoir, of the receptacle and the lines can heat loss be reduced in these areas, resulting in lower energy needs causes the device. In particular, by isolating the Lines the risk of clogging are additionally reduced.

According to one particular embodiment can the device is a temperature-controlled, in particular heatable or coolable third storage vessel to hold on a dyeing solution, in particular a ethidium bromide, which over a line connected to the receptacle is, include. The third vessel is used thereby the admission of a coloring solution, to Example of an ethidium bromide solution, which of the agarose gel composition are still added in the device can. this makes possible it in addition to producing agarose gels with the desired Agarose concentration, already the dye for staining the To implement DNA into the agarose gel composition. Furthermore is advantageously provided by the automated delivery of Dyeing solution too the agarose gel composition contact the user with the mostly carcinogenic dyes avoided.

Preferably have the first storage vessel for recording The agarose has a volume of about 2 l, the second storage vessel for receiving of the buffer has a volume of about 5 l to about 10 l, the receptacle for receiving the agarose gel composition has a volume of about 1 liter and the third storage vessel for uptake the dyeing solution, in particular the ethidium bromide solution a volume of about 10 to 100 ml depending on the concentration the staining solution. The above Volumes are to be understood as exemplary and limit the Device by no means to the specified values. Rather, vary the volumes of the vessels per interpretation the device.

The Vessels and pipes the device according to the invention are made of glass and / or steel and / or stainless steel and / or plastic or a combination of the aforementioned materials produced.

In Advantageously, at least one of the vessels is interchangeably arranged. The connection to the outlets or the lines can be accomplished for example via a bayonet lock. This allows the vessels to refill or be quickly installed and removed for cleaning. It also makes sense the vessels with one Cover to prevent the ingress of contaminants or Prevent water.

Around achieve a precise dosing of the starting materials in the device to be able to is the first storage vessel, the second Storage vessel and / or the third storage vessel and / or the receptacle a weighing device, in particular a weighing sensor assigned. about Such a device or via such sensors, which can communicate with the control and / or regulating unit the weight change registered the respective vessels and the respective valve is activated and closed accordingly become.

According to a further embodiment, a valve or a slide is attached to the first storage vessel comprising agarose, via which the Amount of agarose, which is to be transferred from the first storage vessel into the receptacle, can be dosed. To monitor and determine the amount of agarose, there is a weighing device on this vessel which registers the change in weight. After the required amount of agarose has now exited the first storage vessel, the closing of the valve or the slider is initiated via the control and / or regulating unit.

Also it has proved to be advantageous if in the first Stockpiled stockpiled Agarose with a delivery unit, for example with a screw conveyor from the storage vessel into the Line to the receptacle or directly is conveyed into the receptacle, until the appropriate amount of agarose out of the first storage vessel promoted has been and the conveyor unit on the Control and / or regulating unit is switched off.

advantageously, is the control and regulation unit with the taxable and / or regulable Facilities and the sensors and measuring equipment via electrical or optical lines or wireless e.g. connected via a radio connection. The control and regulating device is arranged centrally or decentrally and capable of handling all components of the device independently to control and regulate.

According to one another embodiment have the tax and / or controllable devices each at least one valve and / or one Pump up. Such a combination makes it possible to remove the material flows from the Storage vessels exactly to dose to an agarose gel composition with those of the user desired To make shares. After opening promote the valves the pumps indicate the respective volume with the corresponding amount Buffer and staining solution off the respective storage vessels in the Receptacle, after which the valves are closed again. The dosage of agarose gel compositions out of the receptacle in the gel chamber can in the same way with the help of a valve and / or a pump done.

In a further embodiment is the first storage vessel, the second Storage vessel, the third Storage vessel and / or the receptacle respectively associated with at least one flow meter. After opening the respective valve measures the flow meter is the volume that exits the vessel. After the required volume has escaped from the vessel, the Outlet through Shut down closed the valve.

It It will be apparent to those skilled in the art that those described above embodiments to understand the exact dosage of the starting materials by way of example are. The invention is not limited to these, but can in many ways be varied.

Around to monitor the supply of agarose, buffer and staining solution is It is also advantageous if the first storage vessel, the second storage vessel, the third Storage vessel and / or the Receptacle respectively at least one level gauge assigned. The level gauge can for example, a level sensor, a magnetostrictive level meter, a bypass level indicator, an ultrasonic level gauge and / or be a deep well probe, monitor the level of the respective vessels and the signals to the control and / or Pass control unit.

In In a further embodiment of the invention, it is advantageous that at least one of the vessels of the device comprises a temperature sensor. With the help of temperature sensors The temperature of the individual agarose gel components can be determined and an unintentional solidification of the agarose in the vessels, respectively the lines by turning on the heaters largely be prevented. examples for Thermometers or temperature sensors are resistance thermometers, Bimetal thermometers, electronic temperature sensors, infrared thermometers and thermocouples.

advantageously, is the first storage vessel, the second Storage vessel, the third Storage vessel and / or the receptacle respectively at least one stirring device, in particular for homogenizing the vessel contents, is assigned. With this you can inhomogeneities and in particular concentration differences of the vessel contents be largely avoided. For example, if a stirrer is in the receptacle arranged, so can the agarose during the solution process with the buffer solution and optionally the ethidium bromide solution to a homogeneous agarose gel composition stirred before they over the outlet in the gel chamber is dispensed.

When stirrer are in the present invention, for example, propeller, inclined blade, Disk, anchor, screw spindle and paddle stirrer or magnetic stirrer in To consider. Also can stirrers be arranged in the second and / or fourth vessel to the solutions before Prepare the agarose gel composition for homogenization.

In order to operate the device according to the invention automatically, the controllable devices such as the pumps and valves are designed controllable and / or adjustable, whereby the substance streams in the device automatically via the control and / or regulating unit, which monitors the flow or the weight changes of the storage vessels, can be controlled and / or regulated. This control and regulation can be further improved by arranging a flow meter for monitoring the exiting volume, at least in the region of a line.

According to the invention the control and / or regulating unit is a unit for controlling the temperature. The temperature in the individual vessels or in the individual pipelines is detected by the temperature sensors and passed on to the control and / or regulating unit. The temperature the individual areas of the device can now via the heaters in such a way controlled and / or regulated that the agarose during the Preparation of the agarose gel composition at temperatures greater than about 80 ° C in their liquid State convicted and the agarose gel composition in this liquid state at temperatures from about 45 ° C up to 60 ° C until their bottling is held in the gel chamber.

in the Contrast this can during a longer one Standstill of the device, for example, during the night or during the Weekend, over the control and / or regulating unit activates a so-called rest program , causing the device to a temperature of about 4 ° C to about 10 ° C, preferred to about 5 ° C chilled for example, to slow down the growth of microorganisms.

Farther controls and regulates the control and / or regulating unit of the device the stirring devices and adjusts the stirring speed or speed of the stirrer the viscosities of present in the vessels liquids at. This is advantageously a foaming in the Prevents vessels.

According to one advantageous embodiment has the device has an internal, programmable memory and / or an operating unit with input devices and / or a display on. about the input devices, for example via a keyboard or individual Switch, lets from the user the desired agarose gel composition program and then automate with the desired levels of agarose, buffer and ethidium bromide solution with the chosen one Assemble volumes.

In The programmable memory can be programs be stored, which allow measurement data of individual Capture, manage and further components of the device the device, such as the pumps, valves, slide and stirrer Taxes. Also can in the programmable memory recipes for making different Be stored volumes and compositions of agarose gels, which is selected by the user can be. On the display can the specifications of the gel selected by the user are displayed and above In addition, the levels of the individual vessels displayed become. Also, it makes sense if the programmable memory a program for sterilizing the device comprises. This Program can be selected by the user and leads a automatic sterilization of the device by slow heating to a temperature of about 180 ° C and keeping this temperature above a period of about 10 hours. This will be on the one hand In addition, microorganisms are largely killed be while Slow heating with a heating rate of about 0.5 ° C per minute Agarose residues in liquid Condition transferred and thus can be removed from the device. Also, the device can be a Timer include, for example, the device on After work or on weekends a so-called sleep mode activated and reactivated at the beginning of work.

Farther it is provided, the device according to the invention, for example before filling the Storage vessels with water or to clean water vapor. This may be possible Residues or Microorganisms are removed.

advantageously, includes the device additionally a table, in particular a leveling table, for example with a dragonfly. This table can be placed under the outlet of the receptacle and serve as an aligned support of a gel chamber.

The The present invention will now be described with reference to exemplary embodiments described in detail. For this, refer to the attached drawings Referenced. The same drawings apply in the individual drawings Reference to the same parts. Show it:

1 shows an embodiment of the device according to the invention for the automated casting of agarose gels.

2 a schematic diagram of the device according to the invention.

1 shows a possible embodiment of the device according to the invention 1 , The device 1 includes three storage vessels 2 . 3 . 4 , in which the starting materials for the agarose gel 19 be stored. The first storage vessel 2 store the Aga rose 20 , the second storage vessel 3 contains a buffer 21 , in particular a TAE buffer with a pH of about 8.0, with which in the first storage vessel 2 stocked agarose 20 is absorbed and diluted. The third storage vessel 5 holds a dye solution in stock, especially an ethidium bromide solution 22 , The three vessels mentioned above are pipes 28 with the receptacle 4 connected. At the outlets of the vessels 2 . 3 . 4 . 5 are valves 7 and at the outlets of the vessels 3 . 4 . 5 pump 8th attached, which via electrical lines 27 (not shown in detail) with the control and / or regulating unit 6 are contacted. At the first storage vessel 2 are weighing sensors 29 attached, over which the weight and thereby the amount of the leaked agarose can be determined.

Used by the user via the input device 14 at the control and / or regulating unit 6 a gel is selected, the device becomes 1 activated. About the control and / or regulating unit 6 now the heaters 24 , the valves 7 and pumps 8th activated. By controlling the valves 7 and pumps 8th Now the appropriate amounts of the starting materials from the storage vessels 2 . 3 . 5 in the receptacle 4 discharged and after the solution and homogenization of the components of the volume flow from the receptacle 4 a gel chamber 18 fed. To determine the temperature during the manufacturing process are in the individual vessels 3 . 4 . 5 and several temperature sensors in the area of the pipelines 10 arranged over which the temperature at the respective locations of the device 1 monitored and to the control and / or regulating unit 6 is passed on.

It has proven to be particularly advantageous if the in the first storage vessel 2 stocked agarose 20 with a conveyor unit (not shown), for example with a screw conveyor from the storage vessel in the line to the receiving vessel 4 or in the receptacle 4 is conveyed until the appropriate amount of agarose has been transported out of the first storage vessel and the conveyor unit is switched off. Subsequently, the valves open at the outlets of the second and fourth vessels 3 . 5 and leave the appropriate volumes of buffer 21 and ethidium bromide solution 22 over pipelines 28 in the receptacle 4 from. Order the agarose in the receptacle 4 to solve, the receptacle 4 over heaters 24 heated to temperatures greater than about 80 ° C and the temperature of the agarose gel composition 25 C kept until its filling in the gel chamber at temperatures of about 45 ° C to about 60 ° C. If necessary, it may also be useful to use the buffer 21 and the ethidium bromide solution 22 preheat.

For homogenization of the agarose gel composition 25 is in the receptacle 4 a stirrer 12 which gives a homogeneous distribution of the individual components of the agarose gel composition 25 causes. After one in the control and / or regulation unit 6 programmed stirring time opens the valve 7 at the outlet of the receptacle 4 and the agarose gel composition 25 gets into the gel chamber 18 on the leveling table 16 is arranged, drained. The control and / or regulating unit 6 This is in contact with the individual controllable units of the device such as the shut-off valves 7 , the pumps 8th , the level sensors 9 , the temperature sensors 10 and the stirrer motors 11 and monitors the process of preparation of the agarose gel composition 25 , If, for example, one of the components runs low, this is shown on the display 15 the control and / or regulating unit 6 displayed.

After the preparation of the agarose gel 19 can in the device 1 now a so-called rest program can be activated and the device 1 with the help of the cooling equipment 24 be cooled to a temperature of about 4 ° C to about 10 ° C, preferably to about 5 ° C.

In 2 is an example of the interconnection of individual components with the control and / or regulating unit 6 shown.

As can be seen from the figure, are on the vessels 2 . 3 . 4 . 5 several units arranged for monitoring. How to monitor the temperature sensors 10 the temperature in the respective vessels, the level gauge 9 the filling level of the respective vessels and send the corresponding measured values via electrical lines 27 to the control and / or regulating unit 6 , In addition, on the vessels 2 . 3 . 4 . 5 valves 7 , Pump 8th and a stirrer 12 and heating and / or cooling facilities 24 arranged, via the control and / or regulating unit 6 can be controlled and / or regulated. At the first storage vessel 2 a weighing sensor is arranged, via which the weight change of this vessel can be registered.

Furthermore, in 2 a pipeline, for example the outlet of the receptacle shown, via a heater 24 is heated. Thus, portions of these conduits in which the agarose gel composition may harden due to temperatures below about 45 ° C may advantageously be heated to prevent curing and consequent blockages.

It will be apparent to those skilled in the art that the two embodiments described above are to be understood as exemplary. The invention is not limited to these, but can be varied in many ways without departing from the spirit of the invention.

1

contraption for the automated production of agarose compositions

2

first Storage jar

3

second Storage jar

4

receptacle

5

third Storage jar

6

Tax- and / or control unit

7

Valve

8th

pump

9

level meter

10

temperature sensor

11

stirrer

12

stirrer

13

agitator shaft

14

operating unit with input devices

15

display

16

table

17

Leveling feet

18

gel chamber

19

agarose gel

20

agarose

21

buffer

22

ethidium bromide

23

management

24

heating and / or cooling device

25

Agarosegelzusammensetzung

26

apparatus

27

electrical management

28

pipeline

29

weighing System

30

Flowmeter

Claims (15)

Device for the automated production of agarose gel compositions ( 1 ), with a first storage vessel ( 2 ) for the provision of agarose ( 20 ), a second storage vessel ( 3 ) to provide a buffer ( 21 ) and a receptacle ( 4 ), wherein the receptacle ( 4 ) a heating device ( 24 ), wherein the first storage vessel ( 2 ) and the second storage vessel ( 3 ) each have a controllable and / or controllable device ( 7 . 8th ) for feeding a defined amount into the receiving vessel ( 4 ), wherein the receptacle ( 4 ) an outlet and a controllable and / or controllable device ( 7 . 8th ) in order to be able to discharge part of the vessel contents therefrom, and with a programmable control and / or regulating unit ( 6 ) with the taxable and / or regulated facilities ( 7 . 8th ) connected is. Contraption ( 1 ) according to claim 1, characterized in that the first storage vessel ( 2 ) and / or second storage vessel ( 3 ) a device for cooling ( 24 ) and / or heating ( 24 ) and / or that the receptacle ( 4 ) a device for cooling ( 24 ) assigned. Contraption ( 1 ) according to claim 1 or 2, characterized in that the heating device ( 24 ) comprises an electric heater, a Peltier heater, a fuel heated heater, a radiant heater, and / or a microwave heater. Contraption ( 1 ) according to claim 2, characterized in that the cooling device ( 29 ) comprises a Peltier cooling, a chiller, a closed-cycle cooling with compressor, a direct cooling and / or an indirect cooling. Device according to one of the preceding claims, characterized in that the first and second storage vessel ( 2 . 3 ) via lines ( 28 ) with the receptacle ( 4 ) are connected. Contraption ( 1 ) according to one of the preceding claims, characterized by a heating device which is designed such that at least portions of the lines ( 28 ) are heated. Contraption ( 1 ) according to claim 1, characterized by a temperature-controllable third storage vessel ( 5 ) for holding a dyeing solution ( 22 ), which via a line ( 28 ) with the receptacle ( 4 ) connected is. Contraption ( 1 ) according to one of the preceding claims, characterized in that at least a portion of the device ( 1 ) is isolated. Contraption ( 1 ) according to one of the preceding claims, characterized in that the first storage vessel ( 2 ), the second storage vessel ( 3 ) and / or the third storage vessel ( 5 ) and / or the receptacle ( 4 ) a weighing device ( 29 ) assigned. Contraption ( 1 ) according to one of the preceding claims, characterized in that the controllable and / or controllable devices ( 7 . 8th ) at least one valve ( 7 ) and / or a pump ( 8th ) exhibit. Contraption ( 1 ) according to one of the preceding claims, characterized in that the first storage vessel ( 2 ), the second storage vessel ( 3 ), the third storage vessel ( 5 ) and / or the receptacle ( 4 ) at least one flow meter ( 30 ) assigned. Contraption ( 1 ) according to one of the preceding claims, characterized in that the first storage vessel ( 2 ), the second storage vessel ( 3 ) the third storage vessel ( 5 ) and / or the receptacle ( 4 ) at least one level gauge ( 9 ) assigned. Contraption ( 1 ) according to one of the preceding claims, characterized in that the first storage vessel ( 2 ), the second storage vessel ( 3 ), the third storage vessel ( 5 ) and / or the receptacle ( 4 ) each at least one stirring device ( 11 ) assigned. Contraption ( 1 ) according to one of the preceding claims, characterized by an operating unit ( 14 ) and / or a display ( 15 ). Contraption ( 1 ) according to one of the preceding claims, characterized in that at least one of the vessels ( 2 . 3 . 4 . 5 ) is arranged interchangeable.