PURIFICATION

FIELD OF THE INVENTION

The present invention relates to a device and method for collecting and purifying liquid samples, generally for biological applications. The present invention relates in particular to a pooling device for collecting biological material or biomolecules contained in liquid samples and for purification of the biological material or biomolecules contained in the liquid samples. The present invention further relates to a device and method for simultaneous pooling and purification of liquids from, for example, well plates.

BACKGROUND

High-throughput biological procedures in the fields of genomic and transcriptomic analysis are gaining in popularity.

Such procedures tend to include steps within their pipeline, where distinct samples are pooled from well plates into a single solution, followed by bulk processing in one single tube.

With the increase in the number of samples needed for parallel processing, the pooling and purification steps become more and more arduous forthe user, since there are no dedicated solutions that combine the two.

While much effort is put into the development and optimization of the multiplexing/demultiplexing aspects of those applications, this particular step remains heavily dependent on manual labor.

A typical example of this situation is when a 96-well or 384-well plate contains molecularly barcoded nucleic acids (e.g. RNA, DNA), which then need to be pooled and purified into one single tube. To achieve this, one needs to collect all samples using, for example, single or multichannel pipettes and transfer them to a different container containing DNA or RNA binding buffer, which is long, laborious and difficult to automate.

After thorough mixing, the new solution can then be gradually flown though by a purification column.

Only then, the eluted solution can then be processed according to the respective analysis.

This manual pooling procedure and this purification procedure, although not high in complexity, increases the time required for the analysis and the great dependence on the human factor potentially introduces inaccuracies and variation. In US patent NO.8602958B1 , one device to facilitate faster sample pooling from a microplate is described. There, a standard microplate is coupled with a receptacle for centrifugation. The liquid from the microplate is collected in the reservoir and can be further utilized. This solution, while being simple and effective, restricts the researcher into collecting the entirety of each well’s volume in the final pool. In many cases only a portion of that is needed forthe analysis. Additionally, this device still requires an additional step in the case of DNA purification, that is transferring the pooled solution into spin columns.

Other proposed solutions involving microfluidic techniques, cannot be applied on standard microplates and require custom equipment that could interfere in other procedures of the high-throughput pipeline.

Therefore, the development of new components to facilitate liquid collection in a way that can be integrated with automation systems whilst eliminating the need of multiple transfers of said liquid solution is desirable to reduce experimental time and increase the accuracy of the procedure.

SUMMARY OF THE INVENTION

The present invention addresses the above-mentioned limitations by providing a biological material pooling device for biological or biomolecule purification according to claim 1, a biological material pooling device including a biological or biomolecule purification element according to claim 13, and a biological material pooling and purification system according to claims 14 and 15.

Other advantageous features can be found in the dependent claims.

The device and method of the present disclosure permits, for example, to simplify and automate as well as speed-up liquid collection from standard microplates in general, and also to eliminate or reduce human related inaccuracies and variations. The device and method of the present disclosure in particular permits to simplify coupling of the collection device with a purification system such as a DNA purification system.

Instead, for example, of the common DNA purification procedures that are performed by centrifugation of spin columns, in the device and method of the present disclosure the DNA binding to, for example, the silica matrix can be achieved utilizing vacuum.

The biological material pooling device for biological or biomolecule purification and the biological material analysis method assuring these advantages is disclosed herein in the present disclosure.

The device and method of the present disclosure can assure the merging of the “sample pooling’’ and “purification’’ steps into a single procedure. This can be achieved, for example, by connecting a purification or spin column used in purification kits to a bottom tube of the device (see for example, Figure 3). After mixing, for example, the samples under investigation with the appropriate volume of a buffer, depending on the used purification kit, the liquid solution can then be dispensed in the cavity of the device of the present disclosure. Then, for example, a vacuum/negative pressure can be applied to the open end of the purification or spin column which will drive the liquid through, for example, the purification element such as silica matrix of the spin column.

In an embodiment, the open end of the purification or spin column can be directly connected to the vacuum-operating waste system of a laboratory.

In another embodiment, the purification or spin column can be connected to a waste box, placed between the device of the present disclosure and a vacuum pump, into which the unwanted flowthrough of the purification can be collected and later discarded.

In yet a different embodiment, the purification element such as silica matrix used, for example, in the DNA purification is integrated in the tube of the device of the present disclosure, eliminating the need for additional purification or spin columns (see, for example, Figure 4).

The above and other objects, features, and advantages of the present invention and the manner of realizing them will become more apparent, and the invention itself will best be understood from a study of the following description with reference to the attached drawings showing some preferred embodiments of the invention.

A BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

Figure 1 shows a perspective or three-dimensional view of one exemplary embodiment of a pooling or collection device of the present disclosure.

Figure 2A shows a side view and Figure 2B shows a top view of one exemplary embodiment of the pooling or collection device of the present disclosure.

Figure 3 shows a cross-sectional schematic and side view of an exemplary pooling and purification system according to the present disclosure including an exemplary pooling or collection device according to the present disclosure, which is fitted or attached in an air-tight fashion on top of a purification column, with a tubular element of a vacuum or aspiration system being attached to a lower section of the purification column, zone A of Figure 3 being shown in an enlarged manner and showing the coupling of the purification column and tubular element of a vacuum or aspiration system.

Figure 4A shows a cross-sectional schematic and a side view of another exemplary embodiment of the pooling or collection device of the present disclosure, in which a purification element is included and fitted directly within a bottom housing or opening of the device. Figure 4B is a top view of the device of Figure 4A.

Figure 5 shows measured comparative purification efficiencies showing that the device of the present disclosure provides a very similar purification yield as compared to manual pooling, while advantageously offering a much more straightforward pooling and purification process.

Figure 6A shows a lower portion of the lower section of the purification column of the system of Figure 3, the lowersection comprising attachments protrusions orteeth for efficient coupling ofthe purification column and the tubular element of a vacuum or aspiration system.

Figure 6B shows a lower portion of the lower section of the device of Figure 4, the lower section comprising attachments protrusions or teeth for efficient coupling to a coupling element, such as a tubular element, of a vacuum or aspiration system.

Figure 7 shows an exemplary pooling and purification system according to the present disclosure including a pooling or collection device (cross-sectional side view) according to any one of the embodiments of the present disclosure, which is attached to a coupling element of a vacuum or aspiration system and a waste collection container or a waste system.

Figure 8 shows a further exemplary embodiment of a pooling device according to the present disclosure.

Herein , identical reference numerals are used, where possible, to designate identical elements that are common to the Figures.

DETAILED DESCRIPTION OF SEVERAL EMBODIMENTS

Figures 1, 2A, 2B, 3, and 4A to 4B, and 5A to 5D show exemplary pooling or collection devices 1 according to the present disclosure.

The pooling or collection device 1 is, for example, a biological material pooling or collection device. The device 1 is, for example, for pooling or collecting a liquid containing a material or substance(s) to be analyzed or processed, such as biological material ora biological substance(s).

The pooling or collection device 1 can be used for purification such as biological or biomolecule purification, for example DNA purification. In some exemplary embodiments, the device 1 is configured or arranged to permit simultaneous pooling and purification. The device 1 can thus be a pooling or collection device and a purification device, for example, a biological material/substance pooling and purification device. The device 1 includes or defines a first or input orifice or aperture 5. The device 1 comprises, for example, a receptacle or vessel 3 including or defining the first orifice 5. The device 1 and receptacle 3 are configured to receive at least one liquid through the first orifice 5. As mentioned, the liquid may contain material or substances, such as one or more biological materials or biological substances to be analyzed or processed. The first orifice 5 comprises or defines an (inner) opening or passage through which the liquid enters into the device 1.

The first orifice 5 is, for example, configured to receive at least a portion of, for example, a 96-channel pipette and the device 1 receives the liquid from the 96-channel pipette.

The device 1 or receptacle 3 also includes or defines a second or guiding orifice 7 located opposite the first orifice 5. The device 1 or receptacle 3 may further include or define a third or exit orifice 9.

The second orifice 7 is located below the first orifice 5 and the third orifice 9 is located below the second orifice 7. The first and second orifices 5, 7 may for example be vertically aligned, or the first, second and third orifices 5, 7, 9 may for example be vertically aligned.

The first, second and third orifices 5, 7, 9 may, for example, be superposed at different vertical or relative levels L1 , L2, L3 (see for example Figure 2A). The levels L1 , L2, L3 extend downwards or in a direction of flow of the liquid.

Each level L1 , L2, L3 may, for example, include solely one orifice. The first orifice 5 may, for example, consists solely of a single orifice or comprises or defines a single opening. The second orifice 7 may, for example, consists solely of a single orifice or comprises or defines a single opening. The third orifice 9 may, for example, consists solely of a single orifice or comprises or defines a single opening. This permits efficient collection of a large quantity of liquid and reduces the risk of liquid being lost via spray or rebounds from surfaces of the device 1.

The third orifice 9 may, for example, be located opposite the first orifice 5 and opposite the second orifice 7.

The second orifice 7 receives the liquid from the first orifice 5 and guides the liquid out of the device 1 or guides the liquid to the third orifice 9 and outside the device 1. Figure 3 shows the direction T of transfer of the liquid out of the device 1.

The second orifice 7 is configured and located between the first and third orifices 5, 9 to guide the at liquid to the third orifice 9 and outside the device 1. The device 1 or receptacle 3 includes one or more side walls or panels 11 extending between the first orifice 5 and the second orifice 7. The side wall 11 may extend directly between the first orifice 5 and the second orifice 7.

The one or more side walls 11 may define the first orifice 5. Alternatively or additionally, the device 1 or receptacle 3 may comprise a ridge, frame or protrusion 12 extending from the side wall 11 and comprising or defining the first orifice 5. The ridge, frame or protrusion 12 may, for example, define an annular shape as shown in Figure 1.

The ridge, frame or protrusion 12 may, for example, extend to define a continuous or non-continuous outer perimeter of the device 1. The ridge, frame or protrusion 12 may, for example, include coupling means such as one or more side apertures (non-shown) and/or protuberances (not shown) configured to couple with a holding apparatus for holding device 1. Such coupling means may alternatively or additionally be defined by the ridge, frame or protrusion 12 defining a greater radial extension than the side wall 11 to delimit an undercut or indentation N (Figure 8).

The side wall 11 may, for example, extend between the first orifice 5 and the third orifice 9 and may, for example, extend directly between the first orifice 5 and the third orifice 9. The device 1 and receptacle 3 may include one side wall 11 or a plurality of interconnected or coupled sides walls 11.

The side wall 11 , or at least an inner surface(s) S1 thereof, may for example be tapered or converge between the first orifice 5 and the second orifice 7. The side wall 11 or at least an inner surface(s) S1 thereof, may for example be tapered entirely or converge entirely between the first orifice 5 and the second orifice 7.

The side wall 11 may also define the second orifice 7, or a portion thereof.

The side wall (or side walls) 11 extends, for example, between the first orifice 5 and the second orifice 7 to define at least one cavity 15 inside the device 1. The cavity 15 defines an inner volume of the device 1 communicating between the first and second orifices 5, 7, the cavity 15 (temporarily) receives the liquid to be analyzed or processed, following passage of the liquid through the first orifice 5.

The cavity 15 comprises or consists of a cavity tapering or converging inwardly, in a direction of a center C of the device 1 (Figure 2A). The cavity 15 tapers or converges inwardly, for example, in a direction extending towards the second orifice 7 or in a direction extending downwards towards the lower level L2 of the device 1. The sidewall 11 , or at least an inner surface(s) S1 thereof, tapers or extends inwardly towards the center C orthe second orifice 7 of the device 1 to define the cavity 15.

The center C is, for example, located at a geometric center or a center of symmetry of the device 1. The side wall 11 , or at least an inner surface(s) S1 thereof, may extend, for example, between the first orifice 5 and the second orifice 7 to define the cavity 15 of decreasing cross-sectional area AR, that decreases in cross-sectional area AR (Figures 1 and 2A) when extending between the first orifice 5 and the second orifice 7. The cross-sectional area AR is, for example, defined as the area of a plane extending (substantially) perpendicular to the center C of the device 1 and between the side wall or side walls 11 , as for example shown in Figure 2A. Figure 2A shows an exemplary cavity 15 where the exemplary area AR2 is less than the area AR1.

The area AR may for example reduce continuously and/or non-continuously when going from level L1 to level L2.

The cavity 15 defined by the side wall 11 , or at least an inner surface(s) S1 thereof, permits to guide the liquid through the receptacle 3 and device 1 and permits to remove the liquid out of device 1.

The side wall or walls 11 (or at least the inner surface(s) S1 thereof) may, for example, extend linearly or non-linearly. The side wall or walls 11 (or at least the inner surface(s) S1 thereof) may, for example, extend to define non-acute or obtuse angles on the inner surface of receptacle 3 or cavity 15 between the first orifice and the second orifice.

The side wall or walls 11 , may for example, define an inner slope percentage of the inner surface S1 of at least 75%, or a slope percentage SL% of the inner surface S1 where 275%>SL%>75%. The slope percentage is defined by (H1/D1) x 100 (see Figure 2A).

The outer slope percentage of the outer surface S2 may be the same as the inner slope percentage.

Alternatively, the at least one side wall 11 defines an angle a with the center of the second office 7 or the center C of the device 1 of at least 35°, or the angle a where 65°>a>35°. The exemplary angle a shown in the embodiment of Figure 2A is approximately 45°.

As mentioned, the device 1 and receptacle 3 are configured to receive the liquid to be analyzed or processed through the first orifice 5, and the cavity 15 is configured to guide the liquid, or to communicate or transfer the liquid from the first orifice 5 to the second orifice 7 to remove or permit the removal of the liquid from the device 1.

The side wall or walls 11 or the at least the inner surface(s) S1 thereof may, for example, comprise or consists solely of chemically inert material and/or hydrophobic material. Alternatively, the inner surface(s) S1 of the side wall or wall 11 may include or consist of a chemically inert material and/or hydrophobic material deposited on the side wall 11 inside the device 1. The side wall or walls 11 or the at least the inner surface(s) S1 thereof may, for example, comprise or consists solely of polypropylene and/or polystyrene.

The device 1 and some or all elements thereof may comprise or consist solely of for example chemically inert material and/or hydrophobic material. The device 1 and some or all elements thereof may comprise or consist solely of for example polypropylene and/or polystyrene.

The first orifice 5 may, for example, define or comprise a rectangular opening or passage through which the liquid is introduced into the device 1.

The device 1 may further include an elongated tube 17 extending from the receptacle 3 or extending from the second orifice 7 away from the receptacle 3, for example, downwards towards the further lower level L3.

The elongated tube 17 may comprise or define the third orifice 9. Liquid exits through the third orifice 9 (directly) outside of the device 1. As mentioned, the third orifice 9 may, for example, be located opposite the first and second orifices 5, 7.

The second orifice 7 and/or the third orifice 9 may, for example, be located at the geometric center or center of symmetry of the device 1. That is, the geometric center or center of symmetry C of the device 1 passes through the second orifice 7 and/or the third orifice 9.

The elongated tube 17 may define part of the second orifice 7, with the receptacle 3 defining another or remaining part of the second orifice 7.

The elongated tube 17 may, for example, be removably attached or permanently attached to the device 1 and to receptacle 3.

The device 1 may include an attachment or coupling permitting a removable attachment or coupling of the elongated tube 17. This can, for example, be implemented via a mechanical coupling such as via complementary inner and outer threads (or vice-versa) respectively on the receptable 3 and the elongated tube 17, or via a form fit or press-fit between the receptable 3 and the elongated tube 17. A leak-tight or air-tight seal may also be included in the mechanical coupling between the receptable 3 and the elongated tube 17. This permits receptacles 3 of different volume size and/or with different inner liquid transfer profiles to be exchanged and attached to the elongated tube 17 to best adapt to different pooling quantities of liquid and to assure faster liquid analysis and processing.

Alternatively, receptable 3 and the elongated tube 17 may be adhesively attached to form a permanently attached structure, or the elongated tube 17 may be integrally formed in one piece with the receptacle 3, and the one or more side walls 11 extend to define the elongated tube 17 and the third orifice 9. The device 1 can thus be integrally formed in one single piece or be a molded piece, or alternatively be composed of a plurality of elements configured to be coupled together.

The second orifice 7 may, for example, define a rectangular, circular, oval, or elliptical opening or passage.

The third orifice 9 may, for example, define a rectangular, circular, oval, or elliptical opening or passage.

The openings defined by the first, second and third orifices 5, 7, 9 may for example be vertically aligned. The openings may, for example, be superposed at the different vertical or relative levels L1, L2, L3. Each level L1, L2, L3 may, for example, include solely one opening or passage.

The shape of the openings or passages of the orifices are, for example, defined as a cross-sectional shape taken in the plane extending (substantially) perpendicular to the center C of the device 1. The exemplary embodiment of Figures 1 and 2Ato 2B shows a rectangular opening of the first orifice 5, and the second and third orifices 7, 9 defining or comprising circular openings.

The cavity 15 may comprise or consist of, for example, a tapered or conical cavity 15 extending between the first and second orifices 5,7 or in the direction towards the elongated tube 17, as shown in the exemplary embodiment of Figures 1 and 2A to 2B.

The cavity 15 may, for example, define a rectangular cross-section (in the plane extending (substantially) perpendicular to the center C of the device 1) for example along a portion of the cavity extending in the direction towards the elongated tube 17. The cavity 15 may alternatively define or have other cross-sectional shapes such as circular, oval, or elliptical.

The cavity 15 may, for example, comprise a plurality of different cross-sectional shapes that change or develops as the cavity extends towards the second orifice 7. The different cross-sectional shapes may, for example, be amongst those mentioned above. For example, an upper portion UP may define a rectangular cross-section and a lower portion may define a circular, oval, or elliptical cross-section. The cavity 15 may include an intermediate section where the rectangular cross-section may (gradually) transform into a circular, oval, or elliptical cross-section as the cavity extends towards the second orifice or lower level L2. This permits, for example, to first collect a large quantity of liquid and then guide the liquid towards exiting the device 1 for easy removal of all the liquid.

The elongated tube 17 comprises or defines a hollow tube. The elongated tube 17 may, for example, have or define an inner elongated passage 19 extending between the second and third orifices 7,9 to permit the liquid to pass therethrough and out of the elongated tube 17. The inner elongated passage 19 may, for example, define or have the same cross-sectional profile as the second orifice 7 and/or third orifice 9. The elongated tube 17 may have or define for example, a rectangular, circular, oval, or elliptical cross-sectional profile.

The elongated tube 17 defines, for example, an outer profile configured to be coupled to or received in an airtight manner inside a further device or element coupled thereto. An exemplary further device is a purification or spin column 21 for biological matter or biomolecule extraction, for example, extraction or purification of nucleic acids (Figure 3). An exemplary element is for example, a coupling extremity of a vacuum tube ortubing 23 that is connected to a lower extremity LE of elongated tube 17 of the device 1 of the embodiment of Figure 4A.

The elongated tube 17 may, for example, be tapered or converge inwards (towards the center C of the device 1) along its direction of elongation in a downward direction or in the direction of transfer T of the liquid out of the device 1. The elongated tube 17 may, for example, be tapered or converge inwards along a portion thereof, for example, a lower portion or lower half of the elongated tube 17. This allows the elongated tube 17 and the device 1 to be coupled to the further device or element and to be received therein (such as a purification column or vacuum tube coupling extremity), and to form an air-tight connection or contact with this further device or element. The elongated tube 17 can thus be partially received inside the further device or element.

The lower extremity LE of the elongated tube 17 may thus define a smaller width w2 than the width w1 of the upper extremity of the elongated tube 17 (Figures 2A, 4A).

The third orifice 9 may, for example, define an opening of identical size or cross-sectional area to that of the second orifice 7, or the third orifice 9 may define a smaller cross-sectional area or smaller opening than the second orifice 7. The third orifice 9 may define an opening that is between 0.4 and 0.9 times or 0.4 and 0.6 times the size or cross-sectional area of the second orifice 7.

Alternatively, the elongated tube 17 be tapered or converge outwards to receive part of the further device inside the elongated tube 17.

The elongated tube 17 or an inner surface SE may comprise or consist of the same material or have the same material properties previously described in relation to the side wall 11.

As previously mentioned, the first orifice 5 may, for example, define or comprise a rectangular opening or passage through which the liquid is introduced into the device 1. The first orifice 5 may define or comprise an opening or passage having a length L and width W (Figure 2B) of a standard microplate as defined by the standard ANSI SLAS 1-2004 (R2012)with Ls=127.76±0.5mm and Ws=85.48±0.5mm, the length and the width being measured (substantially) cross-sectionally to the center C. The length L and width W can, for example, respectively be between 0.95 and 1.1 times a length Ls of the standard microplate and between 0.95 and 1.1 times the width Ws of the standard microplate.

The second orifice 7 defines or comprises a smaller opening or passage than that defined by the first orifice 5 and defines, for example, an opening that is between 0.05 and 0.2 times the size or cross- sectional area of the first orifice 5. For example, the second orifice 7 may define or comprise an opening of diameter between 0.7cm and 2cm for a circular opening, for example 1 cm. The third orifice 9 may, for example, comprise or define an identical opening or slightly smaller opening, for example, between 0.7 and 0.9 times smaller, and may define for example a diameter between 0.5cm and 1.5cm, for example 0.65cm. The cross-sectional area of the second orifice 7 orthe opening defined by the second orifice is for example, between 0.0075 and 0.02 times the cross-sectional area of the second orifice 7 or the opening defined by the second orifice, for example, 0.01 times the cross-sectional area of the second orifice 7 orthe opening defined by the second orifice.

The device 1 can thus, for example, define a funnel-like structure.

Figures 4A and 4B show another embodiment of the device 1 of the present disclosure in which a incudes the biological or biomolecule purification element 27 is included and fitted directly within a bottom housing or chamber 29 of the device 1. Device 1 is thus a pooling and purification device.

The biological or biomolecule purification element 27 comprises or consists of a structure or membrane configured for solid phase extraction of biological material. Exemplary structures or membranes include porous inorganic structures or membranes, for example, a silica matrix or membrane that can be used to bind DNA in the process of DNA purification, or for example, cellulose-based or ion exchange-based matrices (Promega).

The purification element 27 is contained within the cavity or passage section of the elongated tube 17 and in the chamber or housing 29 and the liquid passes through the purification element 27 when traversing the device 1.

The chamber or housing 29 comprises, for example, at least one support wall 31 configured to hold or retain the purification element 27 within the cavity section of the elongated tube 17.

The at least one support wall 31 extends or tapers inwards towards the center of the device 1 (towards the center C) to define a fourth orifice 33 defining an opening or passage of reduced size or reduced cross-sectional area relative to the second orifice 7 and the first orifice 5. The inwardly sloped support wall 31 defines a retaining surface that prevents the purification element 27 from exiting the device 1 through the third orifice 9 while allowing the liquid to pass through an exit the device 1. This permits the purification element 27 to be held or retained within the elongated tube 17 and later recovered for further processing and analysis. The at least one support wall of the elongated tube 17 extends downwards to define the lower extremity LE and the third orifice 9 defining an opening smaller than that of the second orifice 7 (and the first orifice 5) and which may be substantially the same as that of the fourth orifice 33.

The lower extremity LE of the elongated tube 17 is configured to receive or couple to a suction or aspiration element of an aspiration or vacuum creation means, such as a coupling extremity of the vacuum tube or tubing 23 (not shown in Figure 4A). An airtight attachment is assured, for example, by a form fit or press fit between the elongated tube 17 and the vacuum tube or tubing 23.

As shown in Figure 6B, the lower extremity LE of the elongated tube 17 may for example include a coupling section CS comprising a plurality of protrusions and/or recesses 35 for coupling to the coupling extremity of the vacuum tubing 23. This assures an efficient transfer of the liquid out of the device 1 without leaking orwith reduced leaking of the liquid.

Figure 3 shows another embodiment of a system 37 including the above-described device 1 of the present disclosure. The system 37 is, for example, a biological material pooling and purification system 37 including the biological material pooling device 1 and the previously mentioned purification or spin column 21.

The purification or spin column 21 is configured for biological matter or biomolecule extraction. As is known, the purification or spin column 21 for example comprises a hollow elongated housing or column containing the above-mentioned purification element 27 through which the liquid is passed allowing binding of biological material such as nucleic acid or DNA binding in the purification element 27. The purification element 27 is, forexample, located inthe lower portion LP1 ofthe purification orspin column 21.

The elongated tube 17 is received, at least partially, in an air-tight manner inside the upper portion UP1 of the purification or spin column 21. The elongated tube 17 may for example be attached to the purification orspin column 21 in a press-fit or form-fit manner.

The coupling extremity of the vacuum tube or tubing 23 is connected to a lower extremity LE of the lower portion ofthe purification or spin column 21 in an airtight manner. The vacuum tubing 23 is, for example, a suction or aspiration element of an aspiration or vacuum creation means, for example a vacuum system 41.

The vacuum system 41 may include a vacuum pump VP that create a vacuum or aspiration force to force the liquid through the purification element 27 of the purification column 21. The system 41 may forexample include a vacuum manifold MF, a waste collection container 43 or be connected to a waste collection system of a laboratory permitting the liquid to be transferred to the waste collection container 43 or directly to the waste collection system of the laboratory.

As shown in Figure 6A, the lower extremity LE of the purification column 21 may for example include a coupling section CS comprising a plurality of protrusions and/or recesses 35 for coupling to the coupling extremity of the vacuum tube or tubing 23. This assures an efficient transfer of the liquid out of the device 1 without leaking or with reduced leaking of the liquid.

Figure 7 shows the exemplary pooling and purification system 37 including the pooling or collection device of Figures 1 , 2A and 3, the purification or spin column 21 attached to the coupling element of the vacuum tube or tubing 23 of the vacuum or aspiration system 41 , as well as the waste collection container or waste collection system 43 that receives the liquid forced through the purification or spin column 21 by the vacuum or aspiration force.

The device 1 shown in Figure 7 can be replaced by the device 1 of the embodiment of Figure 4A that includes the purification element 27 in the elongated tube 17.

The present disclosure also concerns a method using the device 1 to carry out pooling or pooling and purification of biological matter.

The biological material analysis method includes providing the biological material pooling device 1. The pooling and purification device 1 including the purification element 27 (device of Figure 4A) is attached, for example in an airtight manner, to the vacuum tube or tubing 23 and vacuum system 41, or the pooling device 1 (device of Figure 1 , 2A) is attached, for example in an airtight manner, to the purification column 21 which is attached, for example in an airtight manner, to the vacuum tube or tubing 23 and vacuum system 41. The airtight attachments are assured, for example, by a tight form fit or press fit between the elements being coupled together.

Afterthe addition of a binding buffer solution, such as a DNA or RNA binding buffer, pooling of the liquid containing the biological matter to be analyzed is carried out using the device 1. For example, a 96- channel pipette may be used to transfer the liquid from a multiwell microplate.

A molecular barcoding step ofthe biological material in the wells may also be carried out priorto pooling.

The vacuum system 41 is then used to force the collected liquid in the device 1 through the purification element 27 to carry out a purification step. The liquid transferred through the device 1 can be provided to the waste container 43 or the waste system ofthe laboratory by the vacuum system 41. The biological material collected by the purification element 27 may then be further processed to complete the biological material analysis. For example, washing and elution steps may be carried out depending on the particular analysis being performed.

The above method of the present disclosure may, for example, be carried out as part of genomic and transcriptomic analysis

A use of the described invention is, for example, in the field of transcriptomics and RNA sequencing, where molecular barcoding is quite popular.

For example, approaches for high-throughput RNA sequencing, such as PLATE-seq, BRB-seq and DRUG-seq, have all in common a molecular barcoding step, performed in plates, in which each sample is “tagged” with a molecular barcode during the upstream reverse transcription reaction.

Following barcoding, which can occur in 96-, 384- or 1536-well plates, the generated cDNA samples are normally pooled together by standard liquid handling and purified with one PCR purification column. This process can be trivial for small number of samples but becomes quite challenging for large projects of several hundred samples.

The disclosed invention can be used instead of standard liquid handling to obtain rapid and straightforward pooling of possibly thousands of samples (where for example each sample contains a different “tag”).

The pooler 1 can therefore be used with high-throughput liquid handling solutions and all the wells of SBS well plates can be transferred to the pooler 1 simultaneously, which drastically decrease time and manual effort.

As previously described, a main component of the device 1 is the conical shaped and funnel-like part (Figure 1). The wider opening 5 on top is, for example, designed to receive liquid from up to 96-channel pipettes as its dimensions are well suited for any standard-sized microplate. The bottom narrower opening 9 of the funnel-shaped device 1 can be fitted to most commercially available spin columns 21 in an airtight fashion (Figures 2 and 3), which permits to achieve the vacuum that will drive the flow through, for example, the silica matrix of the spin column 21 towards the waste compartment 43. Alternatively, (Figure 4A), the bottom section of the device 1 can directly contain, for example, the silica matrix that constitutes the purification element 27 of the device 1.

The geometrical features of the funnel-shaped device 1, lacking acute angles and being designed with a steep inner slope, as well as its proposed, but not limiting, material of polypropylene and polystyrene which are chemically inert and maintain a strong hydrophobic behavior ensure that there will be zero to negligible amount of liquid loss on the inner walls of the cavity 15 of the devicel . The following experiment was performed to demonstrate the proof of concept of the proposed invention compared to a typical method of sample pooling, i.e. manual pipetting, using either the Zymo research DNA Clean & Concentrator-5 (cat#D4013) or the Qiagen MinElute PCR Purification Kit (cat #28104).

• Each well of eight 96-well plates (four for Qiagen, four for Zymo) was filled with 15ng of GeneRuler 1 kb DNA ladder (cat # SM0311) in DNA binding buffer.

• 8 spin columns 21 (four from Qiagen, four from Zymo) were placed on a commercial vacuum manifold.

• Manual pooling and purification (slow): four plates were manually pooled by standard pipetting into a falcon tube and then transferred gradually into two columns 21 from Qiagen and two from Zymo.

• Pooling and purification into the pooling/purification device 1 (fast): the device 1 (as shown for example in Figures 1 to 3) was first fitted on the tip of the remaining purification columns 21 (two from Qiagen and two from Zymo). Then four plates were pooled using a 96-tips pipetting head into the device 1 , and the liquid was flown through the purification columns 21 by vacuum force.

Finally, the 8 spin columns 21 were removed from the vacuum manifold and processed as per manufacturer instructions.

The purified DNA was measured by Qubit and the resulting purification efficiencies are reported in Figure 5. As can be seen, the proposed device provides a very similar purification yield as compared to manual pooling, while offering a much more straightforward pooling and purification process.

While the invention has been disclosed with reference to certain preferred embodiments, numerous modifications, alterations, and changes to the described embodiments, and equivalents thereof, are possible without departing from the sphere and scope of the invention. Accordingly, it is intended that the invention not be limited to the described embodiments and be given the broadest reasonable interpretation in accordance with the language of the appended claims. The features of any one of the above-described embodiments may be included in any other embodiment described herein.