IP Library Granted Patent US 7,993,910
Granted Patent B2
US 7,993,910 · App. 12/372,904 · Granted Aug 9, 2011

Methods and apparatus for amplification of DNA using sonic energy

Assignee: DxNA LLC
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Quick Facts
Patent No.
US 7,993,910
App. No.
12/372,904
Granted
Aug 9, 2011
Kind
B2
Abstract

Apparatus and methods for amplification of DNA are provided that use sonic energy in place of conventional thermocyclers. In one embodiment, sonic energy is applied to a PCR cocktail to effect dissociation of double stranded DNA into single strands of DNA. A quiescence stage, where no sonic energy is applied, results in amplification of DNA. Repetition of the cycles of application of sonic energy and cessation of application of sonic energy results in DNA amplification.

Claims (36)

1. A cycler for DNA amplification, comprising:

a structure to hold a sample of material including having double stranded DNA for conversion into single strand DNA for amplification, a polymerase enzyme, nucleosides and primers;

a holder configured to hold said structure, said holder being formed of solid material;

a source of sonic energy in contact with said holder to transmit sonic energy through said holder to said structure and to said material, said source being operative to transmit sonic energy at a frequency, at an energy level and at a duty cycle to dissociate said double stranded DNA into said single stranded DNA without raising the temperature of the double stranded DNA to a temperature at which thermal denaturation will occur;

a controller connected to said source, said controller being configured to supply control signals to said source at a duty cycle

in which said source is first activated to cause said source to generate sonic energy at a frequency and at an energy level and for a time sufficient to effect dissociation of double stranded DNA without raising the temperature of a sample in said structure to cause thermal denaturation thereof, and

in which said source is next deactivated for a period of time selected for amplification of said single stranded DNA.

2. The cycler of claim 1 , further comprising a monitor positioned proximate said structure, said monitor being operative to monitor the status of said amplification.

3. The cycler of claim 2 , wherein said sample of material includes a florescent material, and wherein said monitor includes a fluorescent system positioned to excite said florescent material to cause said florescent material to illuminate in said sample of material, and wherein said florescent system includes a detector configured to measure fluorescent illumination.

4. The cycler of claim 2 , wherein said monitor is a spectrophotometer.

5. The cycler of claim 1 , wherein the said source of sonic energy is a piezoelectric transducer.

6. The cycler of claim 1 , wherein said source of sonic energy is an electromagnetic transducer.

7. The cycler of claim 1 , wherein said source produces sonic energy at or above a frequency of 20 KHz.

8. The cycler of claim 1 , wherein said source has a resonant frequency and wherein said source produces sonic energy at or near the resonance frequency of said source.

9. The cycler of claim 1 , wherein said duty cycle of said controller turns said source on for 2 seconds, and then off for 11 seconds.

10. The cycler of claim 1 , wherein said source operates at a frequency selected to form a standing wave in the said sample of material.

11. The cycler of claim 1 , wherein said duty cycle is selected to effect the dissociation of double stranded DNA and amplification at a substantially isothermal temperature.

12. The cycler of claim 1 , wherein said duty cycle is selected to effect said dissociation of double stranded DNA and said amplification of said single strand DNA is performed without cooling the said mixture.

13. A cycler for DNA amplification, comprising:

a vial containing a reaction mixture including double stranded DNA, a polymerase enzyme, nucleosides and primers;

a vial holder configured to receive said vial, said vial holder being made of solid structure for the transmission of sound therethrough;

a source of sonic energy in physical contact with said vial holder to transmit sonic energy through said vial holder to said vial and to said reaction mixture, said source being operative to transmit sonic energy at a frequency, at an energy level and at a duty cycle to dissociate double stranded DNA into single stranded DNA without raising the temperature of the double stranded DNA to a temperature at which thermal denaturation occurs;

a controller connected to said source, said controller being configured to supply control signals to said source at a duty cycle ,

in which said source is first activated to cause said source to generate sonic energy at a frequency and at an energy level and for a time sufficient to effect dissociation of said double stranded DNA into said single stranded DNA,

in which said source is next deactivated for a period of time selected to allow amplification of said single stranded DNA; and

said duty cycle being repeated to produce a desired amount of said single stranded DNA.

14. The cycler of claim 13 , wherein said controller sends control signals to said source to cause said dissociation of said double stranded DNA and said amplification at a substantially isothermal temperature.

15. The cycler of claim 13 , further comprising a monitor for monitoring the status of said amplification.

16. The cycler of claim 15 , wherein said reaction mixture includes a florescent material, and wherein said monitor further includes a fluorescent material and wherein said monitor is configured to measure fluorescent illumination.

17. The cycler of claim 15 , wherein said monitor is a spectrophotometer.

18. The cycler of claim 13 , wherein the source is a piezoelectric transducer.

19. The cycler of claim 13 , wherein the source is an electromagnetic transducer.

20. The cycler of claim 13 , wherein the source is configured to generate sonic energy at or above 20 KHz.

21. The cycler of claim 13 , wherein the source has resonant frequency and wherein said controller sends control signals to said source to generate sonic energy at or near said resonance frequency of said source.

22. The cycler of claim 13 , wherein said duty cycle of said controller turns on said source for 2 seconds, and then off for 11 seconds.

23. The cycler of claim 13 , wherein said source operates at a frequency to form a standing wave said reaction mixture.

Assignments (5)
RELEASE OF SECURITY INTEREST Recorded Apr 9, 2020
From: DXNA LLC
To: GLORY BIOVENTURES, LLC
Reel/Frame 052360/0131 →
CORRECTION OF NATURE OF CONVEYANCE BY "COLLATERAL ASSIGNMENT" RECORDED AT REEL 024626, FRAME 0744. DXNA HEREBY CONFIRMS THE "COLLATERAL ASSIGNMENT" IS AN ACKNOWLEDGEMENT OF SECURITY INTEREST, WITH DXNA LLC RETAINING OWNERSHIP. Recorded Apr 21, 2011
From: DXNA LLC
To: GLORY BIOVENTURES LLC
Reel/Frame 026165/0670 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 13, 2011
From: ADVANCED MOLECULAR SYSTEMS, LLC
To: DXNA
Reel/Frame 026120/0338 →
COLLATERAL ASSIGNMENT Recorded Jul 1, 2010
From: DXNA LLC
To: GLORY BIOVENTURES LLC
Reel/Frame 024626/0744 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 18, 2009
From: BICKMORE, WILLIAM D.; ROBERTS, DANVERN RAY
To: ADVANCED MOLECULAR SYSTEMS, LLC
Reel/Frame 022273/0913 →
Continuity (3)
Division 11432935 · May 12, 2006
Provisional Application 60680738 · May 13, 2005
Related Publication 20090155893A1 · Jun 18, 2009