CONVECTION POLYMERASE CHAIN REACTION METHOD

• US 20110033899A1
• Filed: 07/11/2008
• Published: 02/10/2011
• Est. Priority Date: 06/14/2007
• Status: Abandoned Application

• Current Assignee: Institut Biokhimii I Genetiki Ufimskogo Nauchnogo Tsentra Ran
• Original Assignee: Institut Biokhimii I Genetiki Ufimskogo Nauchnogo Tsentra Ran
• Inventors: Garafutdinov, Ravil Rinatovich, Magdanov, Eduard Gavisovich, Chemeris, Aleksei Viktorovich, Urmancheev, Said Fedorovich, Chemeris, Dmitry Alekseevich, Lebedev, Yury Anatolievich, Vakhitov, Vener Absatarovich
• Application Number:
  US12/663,248
• Publication Number:
  US 20110033899A1
• Time in Patent Office: Days
• US Class Current: 435/91.2
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 2400/0445 Natural or forced convection
• CPC Class Codes: B01L 2400/0448 Marangoni flow; Thermocapillary effect
• CPC Class Codes: B01L 2400/0472 Diffusion
• CPC Class Codes: B01L 3/5082 Test tubes per se
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: B01L 7/525 with physical movement of samples between temperature zones
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]

Abstract

The invention relates to a method for rapid detection of specific nucleic acid fragments with the aid of a PCR, consisting in amplifying target products in a special DNA thermocycler, provided with a special reaction thermal unit, which makes it possible to obtain, in reaction vessels in the form of standard polypropylene test tubes, a sloping temperature gradient directed at an angle to the direction of gravity. The amplification time is of 1-5 minutes. The inventive method can be recommended for DNA diagnostics, also field conditions, in medicine, veterinary sciences, in sanitary and epidemiological studies for detecting agents of dangerous infections, including potential bio-terrorist attacks, in criminalistics for identifying criminals in the food industry for detecting food products from genetically modified organisms, for testing raw material quality etc.

Claims

1. A method for amplification of specific fragments of nucleic acids with the aid of a polymerase chain reaction, characterized in that due to the effect of accelerated convection occurring as a result of a force of buoyancy caused by a sloping temperature gradient applied to a reaction vessel and oriented at an angle to the direction of action of gravity, cyclic temperature changes that are necessary for multiple sequential implementation of steps of denaturation, annealing and elongation in the reaction mixture due to constant, directed displacement of liquid layers very rapidly occur.

2. The method according to claim 1, characterized in that a reaction vessel is used that is a standard polypropylene test tube.

3. The method according to claim 1, characterized in that registration of the process of amplification of nucleic acids is carried out in the real time mode.

4. The method according to claim 1, characterized in that the application of identical temperatures to corresponding parts of the reaction vessel and the synchronous change thereof instead of the initiation of a temperature gradient provides sufficient homogeneity of the temperature for the whole liquid and if necessary makes it possible to carry out the usual PCR.