Rapid thermal cycling for sample analyses and processing

Abstract

An apparatus for thermal processing nucleic acid in a thermal profile. The apparatus employs a reactor holder for holding reactor(s) each accommodating reaction material containing the nucleic acid. The apparatus includes a first bath; and a second bath, bath mediums in the baths being respectively maintainable at two different temperatures; and a transfer means for allowing the reactor(s) to be in the two baths in a plurality of thermal cycles to alternately attain: a predetermined high target temperature T HT , and a predetermined low target temperature T LT ; and reciprocating means to enable relative reciprocating motion between the holder and at least one bath while the reactor(s) is/are placed in the at least one bath, the relative reciprocating motion being executable by shaking the bath or the holder or both.

What is claimed is: 
     
         1 . An apparatus for thermal processing nucleic acid in a thermal profile, the apparatus employing a reactor holder for holding reactor(s) each accommodating reaction material containing the nucleic acid, the apparatus comprising:
 a first bath; and   a second bath, bath mediums in the baths being respectively maintainable at two different temperatures; and   a transfer means for allowing the reactor(s) to be in the two baths in a plurality of thermal cycles to alternately attain:
 a predetermined high target temperature T HT , and 
 a predetermined low target temperature T LT ; and 
   reciprocating means to enable relative reciprocating motion between the holder and at least one bath while the reactor(s) is/are placed in the at least one bath, the relative reciprocating motion being executable by shaking:
 a) the at least one bath or, 
 b) the holder or, 
 c) the holder and the at least one bath, 
   the reciprocating motion is at a frequency above 0.2 Hz with above 0.5 mm amplitude.   
     
     
         2 . The apparatus according to  claim 1 , further comprising:
 a third bath, wherein the transfer means and the reciprocating means allow the reactor(s) to be in the third bath with the reciprocating motion to attain a predetermined medium target temperature T MT .   
     
     
         3 . The apparatus according to  claim 1 , wherein the reciprocating motion is substantially in horizontal or vertical direction. 
     
     
         4 . The apparatus according to  claim 1 , wherein the reciprocating means reduces speed of the reciprocating motion as the reactor(s) approach the target temperatures. 
     
     
         5 . The apparatus according to  claim 1 , wherein the reciprocating means stops the reciprocating motion during fluorescent imaging of the reactor(s). 
     
     
         6 . The apparatus according to  claim 1 , wherein the reciprocating means continues the reciprocating motion during fluorescent imaging of the reactor(s) when optical means for illuminating the reaction material and collecting the emitted light from the reaction material is moving with the reactor(s). 
     
     
         7 . The apparatus according to  claim 1 , further comprising:
 a reactor guard comprising reactor confining means to partially confine the reactor(s) to prevent the reactor(s) from getting deformed under resistive forces and the T HT  when the reactor(s) is/are received in the bath medium comprising high thermal conductivity powder and during the reciprocating motion, the reactor confining means facing the direction of the reciprocating motion.   
     
     
         8 . The apparatus according to  claim 7 , wherein the reactor guard is made up of materials comprising metal or glass or high temperature plastics or ceramics. 
     
     
         9 . The apparatus according to  claim 7 , wherein the reactor guard is an extension of the reactor holder. 
     
     
         10 . The apparatus according to  claim 1 , further comprising:
 a bottom support means in the bath bottom for supporting the bottom tip(s) of the reactor(s) during the reciprocating motion when the bath medium is powder, for reducing the bending moment on the reactor(s), wherein the reciprocating motion to the reactor(s) is provided by any one of the methods selected from the group consisting of
 a) moving the reactor holder, 
 b) moving the bath bottom, and 
 c) moving the bath bottom and the reactor holder in opposite directions. 
   
     
     
         11 . The apparatus according to  claim 1 , wherein a temperature stabilization is performed at one of the target temperatures. 
     
     
         12 . The apparatus according to  claim 1 , wherein the reciprocating means provides a three-stage shaking of the reactor(s) in the second bath such that a higher speed shaking is followed by a lower speed shaking followed by no shaking for taking fluorescence images. 
     
     
         13 . The apparatus according to  claim 1 , further comprising:
 a sixth bath to contain a liquid or hot air maintainable at 40-80 degree Celsius, wherein at least a portion of the bath wall is transparent to allow transmission of illumination light from a light source and transmission of emitted light from the reactor(s).   
     
     
         14 . The apparatus according to  claim 1 , wherein the bath medium in any of the baths is in at least one phase selected from the group consisting of air, liquid, solid, and powder. 
     
     
         15 . The apparatus according to  claim 1 , further comprising a reactor temperature sensor configured for moving with the reactor holder during thermal cycling, to monitor temperature of the reactor(s). 
     
     
         16 . The apparatus according to  claim 20 , further comprising a vessel containing a substance to encapsulate the reactor temperature sensor, the vessel and the substance having similar construction or heat transfer characteristics to that of the reactor(s) and the reaction material. 
     
     
         17 . The apparatus according to  claim 1 , wherein, at least one bath has a length to width ratio of (2-10):1, the reciprocating motion being conducted along the length direction. 
     
     
         18 . A method for thermal processing of nucleic acid in a thermal profile employing the apparatus according to  claim 1 , the method comprising:
 employing a reactor guard comprising reactor confining means to partially confine the reactor(s) to prevent the reactor(s) from getting deformed under resistive forces and the T HT  when the reactor(s) is/are received in the bath medium comprising high thermal conductivity powder and during the reciprocating motion, the guard facing the direction of the reciprocating motion.   
     
     
         19 . The method according to  claim 18 , wherein the reactor guard is made up of materials comprising metal or glass or high temperature plastics or ceramics. 
     
     
         20 . The method according to  claim 18 , wherein the reactor guard is an extension of the reactor holder. 
     
     
         21 . The method according to  claim 18 , wherein the reactor is in the form of capillary closed at one end. 
     
     
         22 . The apparatus according to  claim 1 , wherein the reciprocating means comprises a reactor shaker for shaking the holder, the shaking being independent of any movement of the holder as provided by the transfer means. 
     
     
         23 . The apparatus according to  claim 1 , wherein the transfer means comprises the reciprocating means and in operation the shaking of the holder is executed by the transfer means. 
     
     
         24 . The apparatus according to  claim 1 , wherein the frequency and the amplitude are selected from the group consisting of:
 i) frequency above 1 Hz with above 0.5 mm amplitude,   ii) frequency above 0.2 Hz with above 5 mm amplitude,   iii) frequency above 1 Hz with above 5 mm amplitude,   iv) frequency above 3 Hz with above 0.5 mm amplitude, and   v) frequency above 3 Hz with above 5 mm amplitude.