Data processing: generic control systems or specific application – Generic control system – apparatus or process – Sampled data system
Reexamination Certificate
2000-08-30
2003-10-14
Gordon, Paul P. (Department: 2121)
Data processing: generic control systems or specific application
Generic control system, apparatus or process
Sampled data system
C700S090000, C700S300000, C702S020000, C435S286100, C435S287200
Reexamination Certificate
active
06633785
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates to a thermal cycler and a DNA amplifier method for amplifying nucleic acid of the DNA.
DESCRIPTION OF THE RELATED ART
In the case of inspecting how nucleic acid (gene) in a gene-recombinated crop influences on the human body or in the case of inspecting gene of a patient, the nucleic acid in the crop or nucleic acid particular to the patient must be extracted from respective individual. However, in order to provide nucleic acid of an amount necessary for inspection, extracted nucleic acid must be amplified and there is PCR (polymerase chain reaction) method as the amplifying method. The PCR method is featured in being highly accurate and highly reliable in order to directly analyze gene with less influence by heat.
According to PCR method known as a method of amplifying efficiently such a small amount of DNA (Deoxyribonucleic acid), one cycle is constituted by a step of denaturing DNA by maintaining a micro tube holding DNA at inside thereof at a temperature of around 95° C., a step of annealing DNA by maintaining DNA at a temperature of around 55° C. and a step of amplifying DNA by maintaining DNA at a temperature around 70° C. and DNA is amplified by repeating the cycle (refer to U.S. Pat. No. 4,683,202). In carrying out the PCR method, it is important to use an apparatus capable of controlling temperature with high accuracy since an efficiency of amplifying DNA is increased by accurately controlling the thermal cycle of the respective steps.
Further, as another amplifying method, there is known NASBA method in which nucleic acid is amplified at a constant temperature of 50 through 60° C.
However, highly accurate temperature control is needed even in NASBA method.
FIG. 1
shows a conventional example of thermal cycler which is an apparatus for automatically carrying out PCR method.
The thermal cycler is provided with a metal block
101
inserted with micro tubes
100
, wells
102
, a heater
103
and a cooling pipe
104
.
The micro tubes
100
including a sample are inserted to the wells
102
engraved to the metal block
101
comprising aluminum and in the metal block
101
, temperature of the micro tubes
100
is controlled by using the heater
103
and the cooling pipe
104
to thereby amplify DNA of the sample.
Normally, the wells
102
are formed at about one hundred portions in the metal block
101
and the micro tubes
100
of about one hundred pieces, are simultaneously processed.
Further, when DNA used for research is amplified, the kind of DNA is previously specified and therefore, an amount of about several microliters is sufficient, however, when unknown DNA used for inspection is amplified, an amount of about several milliliters is needed. Thereby, an enormous time period is taken for amplifying DNA to a desired amount.
However, in the above-described conventional apparatus, all of the micro tubes
100
of about one hundred pieces are simultaneously heated or cooled by the heater
103
and the cooling pipe
104
and therefore, it is difficult to uniformly control temperature. This is because temperature of the inserted micro tubes
100
(sample) is controlled by heating or cooling the metal block
101
inserted with the plurality of micro tubes
100
. Therefore, there is a concern that temperature of the micro tubes
100
becomes nonuniform depending on positions of the metal block
101
and there is a possibility that an amount of product after reaction differs by the respective micro tubes
100
and becomes incomplete.
Further, individually different temperature control cannot be carried out for the respective micro tubes
100
and accordingly, for example, even when one hundred pieces thereof can be processed simultaneously, when the processing is started by inserting only ten pieces of the micro tubes
100
to be processed into the wells
102
, the processing efficiency is lowered. Further, there poses a problem in which when the processing is on standby until one hundred pieces of the micro tubes
100
have been prepared, a time period of processing is increased.
SUMMARY OF THE INVENTION
Hence, the present invention has been carried out in view of the above-described conventional problem and it is an object of the present invention to provide a thermal cycler and a DNA amplifier in which highly accurate temperature control can be carried out with regard to individual micro tubes and the processing efficiency is promoted.
In order to achieve the above-described object, according to an aspect of the present invention, there is provided a thermal cycler comprising a plurality of wells capable of containing micro tubes holding a sample including nucleic acid, a plurality of heaters provided at the respective wells for directly or indirectly heating the micro tubes, a plurality of temperature sensors measuring temperature of the micro tubes, and a control apparatus inputted with measured values of the temperature sensors, supplying current to the plurality of heaters based on the measured values and controlling the temperature of the respective micro tubes independently from each other.
According to another aspect of the present invention, there is provided a thermal cycler comprising a plurality of wells capable of containing micro tubes holding a sample including nucleic acid, a plurality of nozzles provided at the respective wells jetting a medium to the wells, a plurality of heaters provided in the nozzles heating the medium, a plurality of temperature sensors measuring temperature of the micro tubes, and a control apparatus inputted with measured values of the temperature sensors, supplying current to the heaters based on the measured values and controlling the temperature of the respective micro tubes independently from each other.
According to another aspect of the present invention, there is provided a thermal cycler comprising a plurality of cylindrical wells which are capable of containing micro tubes holding a sample including nucleic acid, one end portion of each of which is formed with an opening portion for inserting the micro tube and other end portions of which constitute bottom portions, a plurality of temperature sensors installed in contact with outer walls of the wells measuring temperature of the micro tubes, a plurality of heaters arranged to surround the outer walls of the wells or proximately thereto heating the micro tubes, a case which is a case including a well chamber and an air chamber partitioned by a partition wall and in which the well chamber is arranged to align with the plurality of wells by protruding the opening portions of the wells to an outer side thereof and the outer walls of the wells having the temperature sensors to an inner side thereof and the air chamber includes a plurality of air fans, a plurality of cooling nozzles which are nozzles for cooling the micro tubes by jetting air to the wells, attached to be opposed to the bottom portions of the wells at positions of the partition wall in correspondence with the respective wells jetting air from the air chamber to the wells in the well chamber, and a control apparatus connected to the heaters, supplying current to the heaters in accordance with outputs of the temperature sensors controlling the temperature of the respective micro tubes independently from each other.
According to another aspect of the present invention, there is provided a thermal cycler comprising a plurality of wells capable of containing micro tubes holding a sample including nucleic acid and pasted with an indicator which differs in accordance with the respective sample, a plurality of pick up sensors detecting the indicator, a plurality of heaters provided at the respective wells directly or indirectly heating the micro tubes, a plurality of temperature sensors measuring temperature of the micro tubes, and a control apparatus inputted with measured values of the temperature sensors, supplying current to the heaters based on the measured values and controlling the temperature of the respective micro tubes independently from each other by a prev
Iwasaki Hideo
Kasahara Akihiro
Kawano Koichiro
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