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Temperature Control for PCR Thermo Cycler based on Peltier-Effect

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 March - April 2020
ISSN: 0193-4120 Page No. 14303 - 14308

Temperature Control for PCR Thermo Cycler based

on Peltier-Effect Thermoelectric
Dr.S.P.Selvaraj 1, Logasundari V 2, Kumaresan K 3
1
Professor, 2JRF (DST-IDP Project), 3PG student
1,2,3
Department of Electronics and Instrumentation, Bannari Amman Institute of Technology, Sathyamangalam

Article Info Abstract

Volume 83
Page Number: 14303 - 14308
Publication Issue:
March - April 2020
Article History
Article Received: 24 July 2019
Revised: 12 September 2019
Accepted: 15 February 2020
Publication: 21 April 2020
In this paper, a brief study on repeated thermal cycles of (PCR) Polymerase Chain Reaction
is carried out. The process includes high rate of rise in temperature up to certain limit, hold
there for short time and bring back to certain limits hold and continue. The control is
designed for heat exchanger model in the Polymerase Chain Reaction process in order to
maintain a pre-specifiedtemperature sets for certain time periods at different levels. The heat
exchanger consists of the feed-forward and feedback control; for rejection of disturbances in
measurement feed forward control method helps to a great deal. To improve the dynamic
performance while tracking temperature feedback control serves the purpose well enough.
Heat exchanger model for in PCR is studied with step response around operating regions. To
resultant simulations suggest in improved performance with hybrid control strategy as
compare to PI controller.Improve performances both set-point response and steady state
performance were observed, and hence the accuracy and efficiency of PCR is expected to
improve.
Keywords: Polymerase Chain Reaction, Heat Exchanger, Heat Exchanger

I. INTRODUCTION under observation.During every cycle,the DNA

(PCR) Polymerase chain reaction is a terminology
associated with molecular biology,
This technique is used to amplify a single or a
couple of duplicates of any DNA sample, and are
magnified upto multiple order of their magnified.
And thus it is easily generating thousands to even
thousands and thousands duplicates of a sample
DNA .As of now, PCR is simply to go technique
used significantly in the field of medical and
organic research labs for numerous application. In
pcr technique requires three major steps, via.
Denaturation, Annealing and Extension [4]
.molecular biology emphasize the exponential
amplification of nucleic acid chain, and this process
is considered to be the major initial step for them. In
PCR technique, a cyclic order of heating and cooling
to three specific temperatures are involved,which
result in such huge amplification of the sample
Published by: The Mattingley Publishing Co., Inc.
sample doubles the previous amount. The process is
involving three major steps as: denaturation, which
occurs at around 95℃ of temperature. Then comes
the process of annealing, it occurs at around 55℃ of
temperature and finally the extension that occura at
medium temperature of around 72℃. These
repeating process results in a few billion copies of
the sample DNA in around 30-40 complete
cycle[2].For effective and efficient of PCR process,
the thermal well containing the DNA sample must
change their temperature very quickly, repeatedly
and accurately during its thermal cycling. This is the
primary function of a special thermocycler heavily
rely on peltier-effect for their performance based on
their thermoelectric principle based cooling or
heating.
The peltier element and thermal well combination
result in a large mass as well as these components
have lower thermal conductivity, this hinder the
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heating and cooling rates [3]. Till today, most part of
these thermal cycling for a general thermocycler
were governed and controlled with the help of
ordinary PID controllers because of their simplicity,
good stability and considerable robustness [5]. But
use of a conventional PID control restricts
amplification at ramp rates during those thermal
cycles. In case we want to improve the ramp rates by
compromising the accuracy is temperature, the
whole system accuracy is affected. It is the timing
accuracy and temperature transition speed of PCR
that is most important area of interest for researcher.
Thus it made compulsory to use advanced
temperature control algorithm for desired
performance improvement during those thermal
cycles.
II. LITERATURE SURVEY
Tune Shaojig, Qinqin suggested that the framework
for thermoelectric coolers temperature control
mechanism, which is dependent on versatile NN-
PID [11]. A more dynamic system for them is
derived using little flag linearizationtechnique. For
our current model there are two poles and a zero.
This dynamic thermoelectric cooler model is used in
vacillated distinctive working condition. Normal
direct powerful model for such cooler with strong
temperature control structure is proposed for the
chilly end of it using versatile NN-PID calculation.
A better steady state response and dynamic response
can be achieved with this for a conventional step
response.
Thermoelectric cooler is additionally utilized in
aviation and spaceflight field. Temperature control
of thermoelectric cooler does not depend on just
nonlinear characteristics for thermoelectric module,
but rather additionally heat sink and load of cool
end.
Dr.Mohini Joshi, Dr.J.D. Deshpande proposed a
PCR (Polymerase chainreaction) as a scientific
approach in the field of molecular biology to obtain
multiple duplicate copies of DNA from a singleor a
Published by: The Mattingley Publishing Co., Inc.
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ISSN: 0193-4120 Page No. 14303 - 14308
couple of it .A single piece of DNA can be used for
generating a few thousand to thousands of thousands
duplicates of specific DNA arrangements. PCR is
currently a commonlyused or we can say an
indispensable technique utilized in the field of
organic or medical research labs for numerous
applications [4].Three major steps involved in the
PCRtechnique viz. denaturation, annealing and
extension. PCR is actively used for amplification of
DNA sample for investigation and diagnosis of
numerous disease.PCR can be utilizedto distinguish
human qualities as well as qualities of micro-
organisms andinfections. There is a significant use
of PCR technology in legal science laboratories for
proper identification of desired. Denaturation is
aninitial phase in PCR, comprises of continuous
heating of reaction well for around 20-30 seconds at
94-98℃. During this, DNA samples melts down i.e,
denaturation of the conventional double stranded
DNA breaks the hydrogen bond and turns into two
single-stranded DNA parts. The annealing stage
occurs at a considerably lower temperature of
around 50-60℃. This enables the primes to
hybridize in own specific individualcorrelative
strands, an exceptionally valuable tool in the field of
forensic research.The recently framed DNA strand
of groundwork is then bonded to make
indistinguishable duplicates off the first
templatestrand wanted. Taq (ThermusAquaticus)
polymerase accessible nucleotides as to the finish of
the annealed primers. Thus an extensionhappens for
primers as a result of Taq polymerasehappens at
temperature 72℃ for 5 seconds.
The procedures of denaturation,annealing and
extension summarized as one complete cycle.
Numerous cycles needed to intensify DNA focus to
a large number of copies.
Salmiah A, Hasbullah N F, and Samsuddin N I
proposed aSPAD (Single photon avalanche diode)
type of temperaturesensitive gadget [9]. Indeed,
even little assortment of temperature may result in
insecure execution in quantum efficiency, duty, and
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dim checks. In reasonsunstable temperature affectsabilityand gave for all intents and purposes
variation would result all around poor execution of indistinguishable results.
SPAD. Normally,TEC (thermoelectric cooler)
Ismail, Syahirah K N, Normahira M,Aqmariah S N,
would further be used for cooling of photograph
Firdaus A Z A, Zolkifly M Z proposed a
locator.SPAD is fitted on TEC for it to be kept up at
Thermoelectric cooler with minimal performance
an unfalteringtemperature variation due to variety of
coefficient as compared to regular thermocycler.
encompassing temperature. The
TEC reflects a non linear systemmodel during
frameworkstructured utilizing fluffy rationale with
operation [1].The plant is thermoelectric module
the tool compartment in MatlabSimulinkhaving by
makes closed loop systemto minimize heat
and large execution is given as, consistent state
exchange. PID tuning can be muchfaster in
blunder ±0.56℃ andthe settling time for the yield
obtaining good performance for the TEC time
reproduction is 35.91 sec and it is proportionate
response. Higher ordersystem reduced to second
to±2.8%. Sleuth finds it’s useful –ness in
order system for curve fitting method, there
refrigeration gadget to chill offs the
usingboth critical and under damped. PID controls
SPADtemperature. Versatile NN-PID serves as
developed to TEC about time constrainproblems.
temperature control of TEC resultreaction time of
There using the PID control for reducing the power
10℃ outof 70 sec with fulfilling dynamic and
consumption andreduced time constraint.The
enduringexecution. Fluffy self-versatile PID control
method is proposed is integral of time
of temperature with semiconductor laser result in
multipliedabsolute value of error (ITAE) criteria to
upgraded dynamic reaction and ascends inenduring
determine the coefficient of PIDcontroller.
state accuracy. The overshoot is under 1.6% and
direction timediminished to 2.5s with room III PROPOSED METHOD
temperature is 16℃, this control precisionof ±0.005.
Mendy M E, Kaye S, Sande M V D, Rayco-Solon P,
Waigh P A, Shipton D, Awi D, Snell P, Whittle H
and McConkey S J proposed a Quantitative
persistent PCR toobserving HBV serum infection
heap of infection heap of incessant transporters
[8].HBVe antigen to showed unusual state infection
in the blood. PCR strategydependent on SYBR-
green color used to gauge HBV DNA in serum
andplasma. The basis groupings made and proposed Figure.1: Temperature profile
to increase 98 base combineitems from position 379
to 476 of the genome. Warm cycling was performed The thermo cycling of PCR requires following the
inan ABi5700 grouping disclosure system. occasionally changed temperature Profile exactly
Reproducibility over a wide groundbreakingrange and quickly. Such temperature profile is carried
(1.5 × 103 to 1.5 × 108 copies/mL) and connected inthis paper. Figure1 shows the temperature profile
well witha business measure of r = 0.91 and p < to be tracked for the proposed system.
0.001. HBV DNA was a moretried and true pointer a Conventional PI controllers can barely track the
closeness of infection by HBe antigen which profile quickly andprecisely. The best tuned PI
recognized for 77.0% (161/209) of HBeAg –ve and controller capacities like the short dash line. In
over all HBeAg positivebearers. Procedure had thismanner a hybrid control design, a combination
incredible specificity, reproducibility and
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 March - April 2020
ISSN: 0193-4120 Page No. 14303 - 14308

of feed forward and feedbackcontrol activities, is From the below shown figure3 a graph is plotted for
proposed and portrayed in figure 2. temperature interms of voltage (V) and time
(seconds). From this graph a transfer function
isobtained for further processing.

Figure.2: Diagram of the control strategy

The inclining time is as high as 20s and the
overshoot isn't littleenough,thisis the outcome in Figure. 3: Heat Exchanger Dynamics model
diminished execution of the thermo cyclers. simulation data
Thefeedforward control is in-charge of optimizing,
while the feedback controlsactivity for dismissing The controller output varies the temperature in the
aggravations and disposing of the following blunder set point according toexternal disturbances. This
inconsistent state .Where thermo cyclers of the disturbance is associated with an uncertain
device, feedback controller, andfeedforward delayintroduced in the temperature disturbances.
controller are interconnected, theindividual block When the conventional PI controluse without feed
I/Os are, referenceprofile (r), the servo error (e), forward action, introduces peak overshoot takes
plant input (u), plant yield (y), feedback more to settle. Temperature (℃) in terms
activity(uFB), and feedforward activity (uFF).When ofvoltage(V)
the measured temperature is far fromthe set point, The simulation for PI controller for the TEC process
the On-Off control is introduced, which alters the is shown below in figure
warmth squarewith utmost of incline rates and is
connected to temperature control. Comparedwith the
simulation curve of the first order model, the
dynamics model of theheat block can be viewed
approximately as a first order model.
kp
g s =
τs + 1

uBB , e > 𝜀
u={
uFF + uFB , e ≤ ε
Figure .4: PI Controller
A Combined Feed forward-Feedback Control:
IV SIMULATION OF HEAT EXCHANGER
Feedback control is good for set point tracking in
In this paper heat exchanger is designed with
general, while feedforward control can help with
simulation. The transferfunction obtained from the
rejection of measured disturbances. Next we lookat
reviewed paper is used for this simulation.
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the benefits of combining both schemes.
Figure5shows theimplementation combined feed
forward-feedback control in simulationenvironment
for TEC. The feed forward control is used for,
Figure.5:Combined Feed forward-Feedback
Controls
V. RESULT AND DISCUSSION
A comparative graphical representation for the PI
Controller and HybridController is shown in figure 4
and 5. As a result, Hybrid controller(feed forward
and feedback controller) is more efficient when
compared to thePI controller ie,
Table 1 Comparison of PI and Hybrid Controller
parameters
S.No. Parameters/Controller PI Hybrid
1 Time delay 10 9 validated with the hardware.
2 Peak overshoot 1.23 0.9
3 Setting time 85 64
The time delay in PI controller is 10 seconds
whereas in hybrid controllerthe time delay is 9
seconds.
Peak overshoot in PI controller is 1.23 whereas in
hybrid controller it is0.9.
The settling time in PI controller is 85 seconds
whereas in hybridcontroller it is 64.
VI CONCLUSION
For accurate tracking of reference temperature, a
model basedhybrid control strategy is designed and
Published by: The Mattingley Publishing Co., Inc.
March - April 2020
ISSN: 0193-4120 Page No. 14303 - 14308
simulated results were obtained for thethermo
cycler. Related with the traditional PI controller,
figure 6 shows the response of hybridcontroller is
better in terms of time delay settling time, peak
overshoot toenhance the execution and accuracy of
PCR.
Fig. 6: Graphical comparison of PI Controller
and Hybrid Controller
Particularly, the feed forwardactions boost the
bandwidth of the thermo cycler; therefore, the
commandfollowing response is enhanced. Simulated
results indicate that the capabilityand accuracy of
DNA amplification is upgraded by the hybrid
control strategy. As the future scope the control
algorithm will be implemented in realtime and
REFERENCES
1. A.Z. Ahmad Firdaus, M.Z. Zolkifly, K.N.
Syahirah, M. Normahira, S.N.Aqmariah,
Ismail,” Design and Development of Controller
forThermoelectric Cooler System” 2015 IEEE
International Conference onControl System,
Computing and Engineering, 27 - 29 November
2015.
2. B. J. Huang & C. L. Duang. “System Dynamic
Model and TemperatureControl of a
Thermoelectric Cooler.” International Journal of
Refrigeration23.Pp. 1-5, 2000.
3. Debjani Pala and V. Venkataraman K. Naga
Mohan and H.SharatChandra,VasantNatarajan,”
A power-efficient thermocycler based
14307

oninduction heating for DNA amplification by
polymerase chain reaction” ,Article in Review of
Scientific Instruments • October 2004.
4. Dr.Mohini Joshi, Dr.Deshpande J.D,”
Polymerase Chain Reaction: Methods,Principles
and Application” International Journal of
BiomedicalResearch, pg no 81‐ 97, year 2010
5. H. J. Goldsmid. “Application of Thermoelectric
Cooling,” inThermoelectrics Handbook. New
South Wales, Australia: CRC/ Taylor &Francis,
1995, pp. 617-619.
6. Jianing Yang, Yingjie Liu, Cory B. Rauch,
Randall L. Stevens, Robin H.Liu, Ralf Lenigk
and PiotrGrodzinski,”High sensitivity PCR assay
in plasticmicro reactors”, Lab Chip, pg no179–
187 ,8 November 2002.
7. Jorge Aguiar, Licel Rodriguez, Yamila León,
Freya Freyre, José
AngelSilva,MaríaCaridadMontalvo, Omar Gell,
Regla Estrada, Eduardo Canales,VerenaMuzio,
Gerardo Guillén, Eduardo Pentón and Julio
Cesar Aguilar ,”Hepatitis B Virus DNA
Quantification Using a Cost Effective and
SimpleReal Time PCR in Cuban Carriers”,
Journal of Infectious Diseases andTherapeutics,
2014, Vol. 2, No. 1
8. Maimuna E Mendy, Steve Kaye, Marianne van
der Sande, PuraRayco-Solon, Pauline A Weight,
Deborah Shipton, DorkaAwi, Paul Snell, Hilton
Whittle and Samuel J McConkey ,” Application
of real-time PCR toquantify hepatitis B virus
DNA in chronic carriers in The Gambia”
VirologyJournal, 04 April 2006.
9. NurulIzzatiSamsuddin, NurulFadzlinHasbull H,
Salmiah Ahmad “Fuzzy Logic Based
Temperature Control of Thermoelectric Cooler
(TEC) for Single Photon Avalanche Diode
(SPAD) Application”, 4th International
Conference on Mechatronics (ICOM), 17-19
May 2011.
10. R. Marlow & E. Burke. “Module Design and
Fabrication,” inThermoelectric
Published by: The Mattingley Publishing Co., Inc.
View publication stats
March - April 2020
ISSN: 0193-4120 Page No. 14303 - 14308
Handbook.Dallas, Texas: CRC/ Taylor &
Francis, 1995, pp. 597-607.
11. S. Shaojig, Qinqin. “Temperature Control of
Thermoelectric Cooler Based on Adaptive
NNPID.” IEEE International Conference of
Electrical and Control Engineering.pp. 2245-
2249, 2010.
12. K. S. Chong, K. B. Gan , Sue-Mian
Then,”Development of Thermal CyclerUsing
Proportional-Integral Controller for Polymerase
Chain Reaction”IEEE Engineering in Medicine
and Biology 27th Annual Conference,November
2013.
13. XianboQiu ,Jingqi Yuan,” Temperature Control
for PCRThermocyclersBasedonPeltier-Effect
Thermoelectric”, Proceedings of the2005 IEEE
Engineering in Medicine and Biology 27th
Annual ConferenceShanghai, September 1-4,
2005.
14. Yung-Chiang Chung Li-Wei Lai , Lung-Jieh
Yang, “Comparison ofdifferent metal film
thicknesses of cyclic olefin copolymer–
substratepolymerase chain-reaction chips with
single-side and double-side heaters”Conference
Paper in Journal of Micro/ Nanolithography,
MEMS, andMOEMS ,Jul–Sep 2010/Vol. 9_3.
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