received by the International Bureau on 07 December 2017 (07.12.2017)

1. Apparatus for thermal processing of nucleic acid in a thermal profile, the apparatus employing a reactor holder for holding reactor(s) each accommodating reaction material containing the nucleic acid and the reactor(s) being in any form such as tube(s) or wellplate(s) or chip(s) or cartridge(s), the apparatus comprising:

a first bath; a second bath, bath mediums in the baths being respectively maintainable at two different temperatures where the bath mediums may or may not be part of the apparatus; 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 THT, and

a predetermined low target temperature TLT; 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 reciprocating means operating independent of the transfer means and 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.

2. The apparatus according to claim 1 , wherein the reciprocating means is coupled to the transfer means for shaking the holder.

3. The apparatus according to claim 1, wherein the reciprocating means translates a rotational motion to a substantially linear reciprocating motion.

4. The apparatus according to claim 1 wherein the reciprocating means comprises: a motor (103) with a shaft being mounted on the transfer means; a shaker stage (101) that is mounted on the transfer means; and a scotch yoke (115) including a yoke pin (116) in a cam slot (114) being coupled to the shaft, such that a rotation of the motor (103) causes the shaft and the scotch yoke (115) to rotate, causing the yoke pin (116) in the cam slot (114) to push the shaker stage (101) to move linearly in a reciprocative manner thereby shaking the holder.

5. The apparatus according to claim 1 wherein in operation the relative reciprocating motion can be provided at a frequency above 0.2 Hz with above 1mm amplitude.

6. The apparatus according to claim 1 wherein in operation, the relative reciprocating motion is provided at a frequency up to 6 Hz with amplitude up to 3 mm.

7. The apparatus according to claim 1 further comprising:

a third bath where 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 TMT.

8. The apparatus according to claim 1 wherein the reciprocating motion is substantially in horizontal direction.

9. The apparatus according to claim 1 further comprising:

a fourth bath for an additional process for the reactor(s) before the thermal cycling, the additional process being one from the group consisting:

a) reverse transcription-polymerase chain reaction (RT-PCR),

b) hot start process and

c) isothermal amplification reaction, where the transfer means and the reciprocating means allow the reactor(s) to be in the fourth bath with the reciprocating motion to attain an additional process target temperature T_APT.

10. The apparatus according to claim 1 further comprising:

a reactor guard comprising reactor confining means to partially confine the reactor(s) when held by the holder.

11. The apparatus according to claim 10 wherein the reactor guard is made up of materials comprising metal or glass or high temperature plastics or ceramics.

12. The apparatus according to claim 10 wherein the reactor guard is an extension of the reactor holder.

13 The apparatus according to claim 1 further comprising:

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:

a) moving the reactor holder,

b) moving the bath bottom, and

c) moving the bath bottom and the reactor holder in opposite directions.

14. The apparatus according to claim 1 further comprising:

a fifth bath for a temperature stabilization step in the thermal profile.

15. The apparatus according to claim 1, further comprising:

fluorescence imaging means or electrochemical detection means for analyses of the nucleic acid when the reactor(s) is/are in any of the baths or in air outside the baths.

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16. The apparatus according to claim 1, further comprising:

a sixth bath to contain a liquid or hot air maintainable at 40-80degree C, 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).

17. The apparatus according to claim 1 further comprising a reactor temperature sensor that is capable of moving with the reactor holder during thermal cycling, to monitor temperature of the reactor(s).

18. The apparatus according to claiml further comprising:

a seventh bath that can receive the reactor(s) and be progressively heated while conducting melt curve analysis after the thermal cycling.

19. The apparatus according to claiml further comprising:

altering means for altering temperature in any bath during thermal cycling.

20. The apparatus according to claim 1 having at least one bath having a high-aspect-ratio

shape, the length to width ratio being (2-10): 1, the reciprocating motion being conducted along the length direction.

21. Method for thermal processing of nucleic acid in a thermal profile, employing the apparatus according to claim 1, comprising:

accommodating nucleic acid containing reaction material in reactor(s) in any form such as tube(s) or wellplate(s) or chip(s) or cartridge(s);

allowing the reactor holder to hold the reactor(s);

maintaining bath mediums in the two baths at two different temperatures; and

allowing the transfer means to place the reactor(s) in the two baths in a plurality of thermal cycles to alternately attain:

a predetermined high target temperature THT, and

34 a predetermined low target temperature TLT; and

characterized in that:

providing the relative reciprocating motion at a frequency above 0.2 Hz with above 1mm amplitude.

22. The method according to claim 21 comprising:

stopping the reciprocating motion during fluorescent imaging of the reactor(s).

23. The method according to claim 21 comprising:

allowing an optical means to move with the reactor(s) for illuminating the reaction material; and

collecting the emitted light from the reaction material during the reciprocating motion.

24. Method according to claim 21 comprising:

maintaining the THT in the region 85-99 degree Celsius for denaturation of the nucleic acid; and

maintaining the TLT in the region 45-75 degree Celsius for annealing of primers or probes onto nucleic acid or for primer extension.

25. The method according to claim 21 further comprising:

maintaining the temperature stabilization step at one of the target temperatures.

26. The method according to claim 21 comprising:

wherein the bath medium in any of the baths is in any phase including air, liquid, solid, powder and a mixture of any of these.

27. The method according to claim 21 comprising:

employing a vessel containing a substance to encapsulate a 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.

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28. The method according to claim 21 further comprising:

using metallic powder as bath medium in at least one of the baths;

partially confining the reactor(s) by employing a reactor guard having reactor confining means; and

allowing the confining means to face the direction of the reciprocating motion to prevent the reactor(s) from getting deformed under resistive forces of the bath medium and the target temperature when the reactor(s) is/are received in the at least one of the baths.

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