Claims Analysis
Total Claims: 11
Claim 1
Independent Claim
161 words
A method for amplification of DNA, comprising preparing a reaction mixture consisting of a template DNA, primers of synthetic oligonucleotides, desoxynucleosidetriphosphates, and a buffer medium and carrying out thermocontrolled steps of denaturation of the template DNA, annealing with primers, and completion of the complementary chain, with subsequent repetition of the steps of denaturation, annealing and completion until the process is over, characterized by carrying out the process in a continuous closed-loop system consisting of the following thermocontrolled zones: a DNA fusion zone and a cooling zone ensuring denaturation of the template DNA; a zone for annealing with primers, and a zone for completion of the complementary chain, carrying out circulation of a prepared reaction mixture, which functions as a carrier of DNA molecules which pass during the circulation consecutively through each of said zones to be subjected to denaturation, annealing with primers and completion of the complementary chain, the reaction mixture being circulated until a predetermined amount of DNA is prepared.
Claim 2
Dependent Claim
51 words
A method of claim 1, characterized by the fact that the fusion zone has a temperature from 80 to 95°C, the cooling zone has a temperature from 10 to 30°C, the annealing zone has a temperature from 20 to 60°C, and the completion zone has a temperature from 50 to 75°C.
Claim 3
Independent Claim
37 words
A method of any of claims 1-2, characterized by the fact that the circulation is carried out at a heating rate of a reaction mixture from 0.3 to 0.6°C/s with a cooling rate of 8 to 20°C/s.
Claim 4
Independent Claim
84 words
An apparatus for amplification of DNA, characterized by comprising a closed-loop pipeline (1, 2, 3, 4) adapted to be filled with a fluid reaction mixture and by being provided with four thermostats (5, 6, 7, 8) which are mounted downstream one another along the perimeter of the pipeline to define in the pipelines zones with predetermined temperatures corresponding, in the process sequence, to a fusion zone, to a cooling zone, to an annealing zone, and to a zone for completion of the complementary chain.
Claim 5
Dependent Claim
51 words
An apparatus of claim 4, characterized by the fact that temperature is maintained in the fusion zone between 80 and 95°C, in the cooling zone between 10 and 30°C, in the annealing zone between 20 and 60°C, and in the zone for completion of the complementary chain between 50 and 75°C.
Claim 6
Dependent Claim
39 words
An apparatus of claim 4, characterized by the fact that the pipeline (1, 2, 3, 4) is made up of four series-communicating tubes (1, 2, 3, 4), each tube being placed in respective individual thermostat (5, 6, 7, 8).
Claim 7
Dependent Claim
22 words
An apparatus of claim 6, characterized by the fact that the pipeline is made up of capillary tubes (1, 2, 3, 4).
Claim 8
Dependent Claim
43 words
An apparatus of claim 7, characterized by the fact that the tubes are installed on a common base and are detachably connected to one another for replacement for varying the capacity in accordance with the length of a DNA molecule to be synthesized.
Claim 9
Independent Claim
32 words
An apparatus of any of claims 4 through 8, characterized by the fact that a reaction mixture flow velocity controlled (15) is provided in one of them portions of the closed-loop-loop pipeline.
Claim 10
Dependent Claim
25 words
An apparatus of claim 4, characterized by the fact that the closed-loop-loop pipeline is provided with a pump for positive pumping of a reaction mixture.
Claim 11
Dependent Claim
37 words
An apparatus of claim 4, characterized by the fact that the pipeline has a port (16) adapted for provided a liquid stopper, them port being also used for loading a reaction mixture for discharging the end product.