Process and apparatus for fragmenting biomaterials

• US 5,506,100 A
• Filed: 06/01/1993
• Issued: 04/09/1996
• Est. Priority Date: 10/11/1990
• Status: Expired due to Term

• Current Assignee: Indiana University Research & Technology Corporation (Indiana University)
• Original Assignee: Indiana University Foundation (Indiana University)
• Inventors: Kityama, Masahiko, Surzycki, Stefan, Togasaki, Robert K.
• Primary Examiner(s): Wityshyn, Michael G.
• Assistant Examiner(s): LEITH, NANCY J
• Application Number:
  US08/069,580
• Time in Patent Office: 1,043 Days
• Field of Search: 435/6, 435/287, 422/99, 422/100, 422/101, 422/102, 422/243, 422/244, 536/22.1, 239/338, 935/19
• US Class Current: 435/6.12
• CPC Class Codes: B05B 7/0012 Apparatus for achieving spraying before discharge from the apparatus
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: G01N 1/286 involving mechanical work, e.g. chopping, disintegrating, compacting, homogenising microtomes G01N1/06; pulverising in general B02C; mixing in general B01F
• CPC Class Codes: G01N 2001/4066 using difference of solubility between liquid and gas, e.g. bubbling, scrubbing or sparging

Abstract

Described is an improved process for fragmenting a biomaterial and isolating and recovering a component thereof. The preferred improved process includes the step of performing the fragmentation by nebulizing a liquid medium containing the biomaterial. A preferred process for fragmenting isolated DNA includes the step of nebulizing a fluid containing the DNA. This preferred process provides randomness superior to prior known DNA fragmentation methods, as well as other important advantages. Improved nebulization devices are also described.

Claims

1. In a device for nebulization of liquid which has a liquid input and a liquid output, the improvement wherein said device also comprises means for returning substantially all of the nebulized liquid from the output back to the input.

2. A device according to claim 1, and further comprising a barrier spaced from the output for reducing liquid droplets exiting the output to smaller droplets, and means for adjusting the distance of the barrier from the output.

3. The device of claim 1, which includes a nebulization barrier for contacting liquid exiting the liquid output, said nebulization barrier being positionable to selected distances spaced from said liquid output.

4. A device for nebulizing a liquid, comprising:

a base member having internal channels adapted to deliver a liquid sample mist out of an opening in the base member;

an upper member removably connected to the base member and defining an internal chamber into which said mist is delivered;

a nebulization barrier movably attached to the upper member so as to be positionable to selected locations within said chamber so as to be contacted by the liquid sample mist at varying distances from said opening; and

positioning means located externally of said chamber for positioning said nebulization barrier to said selected locations.

5. The device of claim 4 wherein said upper member comprises:

a cylindrical member removably connected to said base member; and

a top member removably connected to said cylindrical member.

6. The device of claim 5 wherein said nebulization barrier is movably connected to said top member so as to be positionable at varying distances from said opening.

7. The device of claim 6 also comprising a sample reservior member removably connected to said base member.

8. A device for nebulizing a liquid, comprising:

a base member having internal channels adapted to deliver a liquid sample mist out of an opening in the base member;

a first upper member removably connected to the base member and defining a first internal chamber;

a second upper member removably connected to the base member and housed within said first internal chamber;

said second upper member defining a second internal chamber smaller in volume than said first internal chamber and into which said mist is delivered;

said second upper member also having an opening; and

a nebulization barrier attached to the first upper member and positionable through the opening in said second upper member to selected locations within said second internal chamber so as to be contacted by the liquid sample mist at varying distances from the opening in the base member.

9. The device of claim 8 wherein said first upper member comprises:

a cylindrical member removably connected to said base member; and

a top member removably connected to said cylindrical member.

10. The device of claim 9 wherein said nebulization barrier is movably connected to said top member so as to be positionable at varying distances from said opening in the base member.

11. The device of claim 10 wherein said base member has liquid input and output openings within said second chamber so as to provide for continuous nebulization of the liquid sample.

12. The device of claim 11 wherein said second upper member comprises:

a cylindrical member removably connected to said base member; and

a conical member removably connected to the top of said cylindrical member, the conical member having an aperture therein and thereby providing said opening in said second upper member.

13. The device of claim 12 wherein said cylindrical member has converging inner walls defining a conical reservior to hold sample liquid.