Rapid thermal cycling for sample analyses and processing

Abstract

An apparatus for thermal processing of nucleic acid in a thermal profile. The apparatus employs a reactor holder for holding reactors to accommodate reaction material containing nucleic acid. The apparatus includes at least two baths separated by thermally insulating partition plate(s) where bath mediums are each maintainable at a predetermined temperature; and a transfer means for allowing the reactors to change position once or plurality of times between any two adjacent baths by selectively opening the partition plate(s) and without lifting the reactors out of the baths.

What is claimed is: 
     
         1 . An apparatus for thermal processing of nucleic acid in a thermal profile, the apparatus employing a reactor holder for holding reactor(s) to accommodate reaction material containing nucleic acid, the apparatus comprising:
 at least two baths separated by thermally insulating partition plate(s) where bath mediums in use are each maintainable at a predetermined temperature; and   a transfer means for allowing the reactor(s) to change position once or plurality of times between any two adjacent baths of the at least two baths by selectively opening the partition plate(s) and without lifting the reactor(s) out of the baths.   
     
     
         2 . The apparatus according to  claim 1 , wherein the at least two baths comprise:
 a first bath where the reactor(s) is/are allowed to attain a predetermined high target temperature T HT , wherein the T HT  is in the region 85-99 degree Celsius for pre-denaturation and denaturation of the nucleic acid; and   a second bath where the reactor(s) is/are allowed to attain a predetermined low target temperature T LT , wherein the T LT  is in the region 45-75 degree Celsius for annealing of primers or probes onto nucleic acid or for primer extension,   the first and the second baths being for thermal cycling the reactor(s) to attain polymerase chain reaction (PCR) amplification or primer extension.   
     
     
         3 . The apparatus according to  claim 2 , further comprising:
 a third bath where the reactor(s) is/are allowed to attain a predetermined medium target temperature T MT , wherein the T MT  is for annealing of primers or probes onto nucleic acid.   
     
     
         4 . The apparatus according to  claim 2 , further comprising:
 a fourth bath where the reactor(s) is/are allowed to attain a predetermined medium target temperature T MT , wherein the T MT  is for extension of primers on nucleic acid.   
     
     
         5 . The apparatus according to  claim 2 , further comprising:
 a fifth bath where the reactor(s) is/are allowed to attain a temperature T AP  for an additional process for the reactor(s) before thermal cycling, the additional process being one selected from the group consisting of
 a) reverse transcription-polymerase chain reaction (RT-PCR), 
 b) hot start process, and 
 c) isothermal amplification reaction. 
   
     
     
         6 . The apparatus according to  claim 2 , further comprising:
 a sixth bath that can be progressively heated while conducting melt curve analysis after thermal cycling.   
     
     
         7 . The apparatus according to  claim 6 , wherein the reactor(s) is/are allowed to stabilize in the first bath, the second bath, the third bath, the fourth bath, the fifth bath or the sixth bath. 
     
     
         8 . A method of using the apparatus of  claim 3 , the method comprising:
 setting the T LT  equal to the T MT .   
     
     
         9 . A method of using the apparatus of  claim 1 , the method comprising:
 using the bath medium in at least one phase selected from the group consisting of air, liquid, solid, and powder.   
     
     
         10 . The method according to  claim 9 , further comprising:
 orienting the reactor(s) held by the reactor holder to allow minimum surface area of the reactor(s) to be in a direction of movement of the reactor(s) between the baths.   
     
     
         11 . A method of using the apparatus of  claim 4 , the method comprising:
 setting the T LT  equal to the T MT .   
     
     
         12 . The apparatus according to  claim 1 , wherein in operation, the partition plates open along a horizontal direction. 
     
     
         13 . The apparatus according to  claim 1 , wherein in operation the partition plates open along a vertical direction.