Low-Cost Temperature Logger for a Polymerase Chain Reaction Thermal Cycler

A thermal cycler is used to amplify segments of DNA using the polymerase chain reaction (PCR). It is an instrument that requires precise temperature control and rapid temperature changes for certain experimental protocols. However, the commercial thermal cyclers are still bulky, expensive and limited for laboratory use only. As such it is difficult for on-site molecular screening and diagnostics. In this work, a portable and low cost thermal cycler was designed and developed. The thermal cycler block was designed to fit six microcentrifuge tubes. A Proportional-Integral temperature controller was used to control the thermal cycler block temperature. The results showed that the maximum temperature ramp rate of the developed thermal cycler was 5.5 °C/s. The proportional gain (Kp) and integral gain (Ki) of the PI controller were 15 A/V and 1.8 A/Vs respectively. Finally, the developed thermal cycler successfully amplified six DNA samples at the expected molecular weight of 150 base pair. It has been validated using the Eppendorf Mastercycler nexus gradient system and gel electrophoresis analysis

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A Prototype Continuous Flow Polymerase Chain Reaction LTCC Device

Materials Science Forum ◽

10.4028/www.scientific.net/msf.539-543.523 ◽

2007 ◽

Vol 539-543 ◽

pp. 523-528 ◽

Cited By ~ 3

Author(s):

Korey Moeller ◽

Jason Besecker ◽

Greg Hampikian ◽

A. Moll ◽

D. Plumlee ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Low Temperature ◽

Continuous Flow ◽

Low Cost ◽

Flow Characteristics ◽

Chain Reaction ◽

Wide Range ◽

Polymerase Chain ◽

Thermal Cycler ◽

Biological Entities

There is a growing need for remote biological sensing in both laboratory and harsh field environments. Sensing and detection of biological entities such as anthrax, Ebola and other micro-organisms of interest involves sampling of the environment, amplification, analysis and identification of the target DNA. A key component of such a sensor is a low cost, portable, reusable, continuous flow polymerase chain reaction (PCR) thermal cycler. Fabrication with low temperature co-fired ceramics (LTCC) can provide a reusable low cost device capable of operating in a wide range of environments The design and manufacture of a prototype continuous flow micro-fluidic PCR device using low temperature co-fired ceramic is presented. Initial modeling of flow characteristics and heat transfer was carried out in SolidWorks™. The prototype device employs resistance heaters below the channels, buried and surface thermocouples for temperature monitoring, and air gaps for thermal isolation.

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DEVELOPMENT OF POLYMERASE CHAIN REACTION THERMAL CYCLER USING NUMERICAL HEAT FLOW ANALYSIS

Journal of computational fluids engineering ◽

10.6112/kscfe.2020.25.4.001 ◽

2020 ◽

Vol 25 (4) ◽

pp. 1-7

Author(s):

Jungseok Kang ◽

Siyoung Yang ◽

Wonchang Cho ◽

Hyung Min Kim

Keyword(s):

Polymerase Chain Reaction ◽

Heat Flow ◽

Flow Analysis ◽

Chain Reaction ◽

Polymerase Chain ◽

Thermal Cycler

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Intracellular amplification of proviral DNA in tissue sections using the polymerase chain reaction.

Journal of Histochemistry & Cytochemistry ◽

10.1177/40.3.1313061 ◽

1992 ◽

Vol 40 (3) ◽

pp. 333-341 ◽

Cited By ~ 67

Author(s):

K P Chiu ◽

S H Cohen ◽

D W Morris ◽

G W Jordan

Keyword(s):

Polymerase Chain Reaction ◽

Cultured Cells ◽

Chain Reaction ◽

Tissue Sections ◽

Mouse Mammary Tumor ◽

Tumor Virus ◽

Polymerase Chain ◽

Thermal Cycler ◽

Cellular Dna

We developed a new method to amplify cell DNA in situ using the polymerase chain reaction (PCR). Proviral sequences of mouse mammary tumor virus (MMTV) contained in cultured cells and tissue sections were amplified intracellularly using a thermal cycler. Two techniques were employed to maintain the localization of the amplified DNA. First, complementary tails at the 5' ends of the oligonucleotide primers resulted in the synthesis of high molecular weight concatamers containing the target sequences. Second, the PCR was carried out in a thin film of agarose solidified over the tissue sections. The specifically amplified and localized DNA was then detected by in situ hybridization (ISH). Our results demonstrate that (a) DNA in tissue sections can serve as the target for the polymerase chain reaction in situ, (b) cell morphology is maintained, and (c) a target of 167 BP can be specifically detected in individual cells. This technique should be generally applicable to amplifying cellular DNA targets in tissue sections for detection in situ.

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An Innovative Rapid Thermal Cycling Device for Polymerase Chain Reaction

Journal of Medical Devices ◽

10.1115/1.2934478 ◽

2008 ◽

Vol 2 (2) ◽

Author(s):

Shadi Mahjoob ◽

Kambiz Vafai

Keyword(s):

Heat Transfer ◽

Polymerase Chain Reaction ◽

Temperature Distribution ◽

Thermal Cycling ◽

Thermal Management ◽

Uniform Temperature ◽

Chain Reaction ◽

Temperature Ramp ◽

Polymerase Chain ◽

Thermal Cycler

Polymerase chain reaction (PCR) is the most commonly used molecular biology technique to amplify nucleic acid (DNA and RNA) in vitro. This technique is highly temperature sensitive and thermal management has an important role in PCR operation in reaching the required temperature set points at each step of the process (denaturing, annealing and elongation). In this work, an innovative microfluidic PCR thermal cycling device is designed to increase the heating∕cooling thermal cycling speed while maintaining a uniform temperature distribution throughout the substrate containing the aqueous nucleic acid sample. The device design is incorporating the jet impingement and micro-channel thermal management technologies utilizing a properly arranged configuration filled with a porous medium. Porous Inserts are attractive choices in heat transfer augmentation. They provide a very large surface area for a given volume which is a key parameter in heat transfer processes. Various effective parameters that are relevant in optimizing this flexible thermal cycler are investigated such as thermal cycler configuration, thickness of inlet and exit fluid channels, fluid flow rate and velocity, the porous matrix material and properties, and utilization of thermal grease. An optimized case is established based on the effects of the cited parameters on the temperature ramp, temperature distribution and the required power for circulating the fluid in the thermal cycler. The results indicate that the heating∕cooling temperature ramp (temperature change per heating∕cooling cycling time) of the proposed device is considerably higher (150.82◻C∕s) than those in literature. In addition, the proposed PCR offers a very uniform temperature in the substrate while utilizing a low power.

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Immuno-capture PCR method for detecting Acidovorax avenae subsp. citrulli from watermelon

Chinese Journal of Agricultural Biotechnology ◽

10.1017/s1479236207001465 ◽

2007 ◽

Vol 4 (2) ◽

pp. 173-179 ◽

Cited By ~ 6

Author(s):

Wang Xiao ◽

Zhang Le ◽

Xu Fu-Shou ◽

Zhao Li-Han ◽

Xie Guan-Lin

Keyword(s):

Polymerase Chain Reaction ◽

Low Cost ◽

Chain Reaction ◽

Direct Pcr ◽

Causal Organism ◽

Bacterial Fruit Blotch ◽

Pcr Method ◽

Polymerase Chain ◽

Honeydew Melon ◽

Melon Seeds

AbstractAn immuno-capture polymerase chain reaction (IC-PCR) method for detection of Acidovorax avenae subsp. citrulli (AAC), the causal organism of bacterial fruit blotch (BFB) of watermelon, was developed by combining the immunosorbent enrichment (ISE) method with classical PCR and comparing with the direct PCR and growth check methods. The results showed that all A. avenae subsp. citrulli strains tested have produced 360 bp specific fragments using IC-PCR and direct PCR methods, while other strains from 10 different genera showed negative PCR results. The minimum detection concentration was about 50–100 cfu/ml and 104 cfu/ml, respectively. The IC-PCR sensitivity was 100 times higher than that of direct PCR. The examination of seven batches of different melon seeds from the markets by IC-PCR showed that one cantaloupe, two honeydew melon and two watermelon seed varieties carried the pathogen, indicating that the IC-PCR is an accurate, sensitive, rapid and low-cost technique.

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Rapid microfluidic thermal cycler for polymerase chain reaction nucleic acid amplification

International Journal of Heat and Mass Transfer ◽

10.1016/j.ijheatmasstransfer.2007.11.014 ◽

2008 ◽

Vol 51 (9-10) ◽

pp. 2109-2122 ◽

Cited By ~ 24

Author(s):

Shadi Mahjoob ◽

Kambiz Vafai ◽

N. Reginald Beer

Keyword(s):

Polymerase Chain Reaction ◽

Nucleic Acid ◽

Nucleic Acid Amplification ◽

Chain Reaction ◽

Polymerase Chain ◽

Thermal Cycler

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Specific Detection of Clostridium botulinum Types A, B, E, and F Using the Polymerase Chain Reaction

Journal of AOAC International ◽

10.1093/jaoac/85.5.1025 ◽

2002 ◽

Vol 85 (5) ◽

pp. 1025-1028 ◽

Cited By ~ 8

Author(s):

Kathy E Craven ◽

Joseph L Ferreira ◽

Mark A Harrison ◽

Paul Edmonds

Keyword(s):

Polymerase Chain Reaction ◽

Clostridium Botulinum ◽

Extraction Procedure ◽

Toxin Gene ◽

Chain Reaction ◽

Gene Target ◽

Human Illness ◽

Polymerase Chain ◽

Clostridial Species ◽

Thermal Cycler

Abstract Clostridium botulinum organisms generally produce 1 of 4 neurotoxin types (A, B, E, and F) associated with human illness. Neurotoxin type determination is important in identification of the bacterium. A polymerase chain reaction (PCR) method was developed to identify 24 h botulinal cultures as potential types A, B, E, and F neurotoxin producers as well as other clostridial species which also produce neurotoxins. Components of the PCR and amplification conditions were adjusted for optimal amplification of toxin gene target regions to enable simultaneous testing for types A, B, E, and F in separate tubes using a single thermal cycler. Each primer set was specific for its corresponding toxin type. A DNA extraction procedure was also included to remove inhibitory substances that may affect amplification. This procedure is rapid, sensitive, and specific for identification of toxigenic C. botulinum.

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