Biochemical amplification of nucleic acids

• US 7,247,429 B2
• Filed: 08/07/2002
• Issued: 07/24/2007
• Est. Priority Date: 08/07/2002
• Status: Expired due to Fees

• Current Assignee: Genetix Ltd.
• Original Assignee: Genetix Ltd.
• Inventors: Hallam, Alysia, Atkinson, George Robert
• Primary Examiner(s): Kim; Young J.
• Application Number:
  US10/212,732
• Publication Number:
  US 20040029235A1
• Time in Patent Office: 1,812 Days
• Field of Search: 435/6, 435/91.1, 435/91.2
• US Class Current: 435/6.11
• CPC Class Codes: B01L 13/02 for receptacle or instruments
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B08B 7/005 by infrared radiation
• CPC Class Codes: B08B 7/0057 by ultraviolet radiation
• CPC Class Codes: C12Q 1/6848 characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
• CPC Class Codes: C12Q 2523/313 Irradiation, e.g. UV irradiation
• CPC Class Codes: G01N 2035/00277 Special precautions to avoid contamination (e.g. enclosures, glove- boxes, sealed sample carriers, disposal of contaminated material)
• CPC Class Codes: G01N 35/1004 Cleaning sample transfer devices

Abstract

A method of cleaning a pin head or other implements between handling a plurality of samples of nucleic acid subject to an amplification process using a biochemical amplification product. The method comprises exposing the pin head or other implement to ultraviolet (UV) and preferably also infrared (IR) illumination between handling different samples in order to suppress cross-contamination by non-specific amplification. The biochemical amplification product may be obtained from a rolling circle amplification (RCA) procedure, such as that catalyzed by bacteriophage phi29, or a polymerase chain reaction (PCR) amplification product.

Claims

1. A method of cleaning an implement between handling a plurality of samples or a plurality of batches of samples of nucleic acid subject to a nucleic acid biochemical amplification process, wherein

the implement comprises a plurality of pins for picking nucleic acid samples, and wherein

the method comprises exposing the pins to illumination with radiation having an ultraviolet (UV) component between handling different samples or batches of the samples, wherein the exposure to the illumination is for a period of between about 4 and 30 seconds and is sufficient to suppress cross-contamination by non-specific amplification.

2. The method of claim 1, wherein the biochemical amplification process comprises rolling circle amplification (RCA).

3. The method of claim 2, wherein the RCA uses a bacteriophage enzyme.

4. The method of claim 3, wherein the bacteriophage enzyme is phi29 polymerase.

5. The method of claim 1, wherein the biochemical amplification process comprises polymerase chain reaction (PCR) amplification.

6. The method of claim 5, wherein the PCR amplification uses thermostable DNA polymerase.

7. The method of claim 6, wherein the thermostable DNA polymerase is Taq polymerase.

8. The method of claim 1, wherein the illumination further includes an infrared (IR) component.

9. The method of claim 1, wherein the illumination is provided by at least one halogen lamp.

10. The method of claim 1, further comprising subjecting the implement to a gas flow to promote cooling during and/or after the illumination.

11. The method of claim 1, further comprising washing the implement.

12. The method of claim 11, wherein the implement is washed in at least one of ethanol and a bleach/water mix.

13. The method of claim 1, wherein exposure to the ultraviolet (UV) component of the illumination inactivates the nucleic acid.

14. The method of claim 1, wherein exposure to the ultraviolet (UV) component of the illumination causes formation of photoproducts which prevent replication of the nucleic acid.

15. A method of using an implement to handle a plurality of samples or a plurality of batches of samples of nucleic acid subject to a nucleic acid biochemical amplification process, wherein the implement comprises a plurality of pins for picking nucleic acid samples, and wherein the method comprises:

(a) picking a first sample of nucleic acid with a pin of the implement, or a first batch of samples of nucleic acid with pins of the implement;

(b) exposing the pins to illumination with radiation having an ultraviolet (UV) component for a period of between about 4 and 30 seconds; and

(c) picking a second sample of nucleic acid with a pin of the implement, or a batch of samples of nucleic acid with pins of the implement.