scholarly journals Pengenalan Teknik Molekuler Polymerase Chain Reaction (PCR) pada Guru Biologi SMA Di Kota Mataram

Jurnal PEPADU ◽  
2020 ◽  
Vol 1 (2) ◽  
pp. 139-143
Author(s):  
Eustachius Hagni Wardoyo ◽  
Yunita Sabrina ◽  
Dewi Suryani
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Hands On ◽  
Post Test ◽  
Polymerase Chain ◽  
Thermal Cycler

Teknik molekuler polymerase chain reaction (PCR) merupakan teknik dasar amplifikasi materi genetik dan protein yang sangat bermanfaat dalam bidang biologi, kedokteran, pertanian dan bidang lain. PCR digunakan dalam identifikasi kelainan genetik, diagnosis mikrobiologi, sekuensing DNA, identifikasi forensik dan masih banyak lagi. Dasar dasar biomolekuler dan teknik PCR ini tertuang dalam kurikulum SMA untuk siswa kelas 3 SMA. Melihat adanya kesenjangan belum terpaparnya semua guru SMA di kota Mataram dengan teknik PCR ini maka perlu dilakukan pengenalan teknik PCR bagi guru SMA dengan harapan akan membantu guru dalam mengimplementasikan materi ini ke dalam pelajaran biologi SMA. Dalam pelatihan ini guru biologi sebagai salah satu perantaraan pembelajaran utama siswa SMA diperkenalkan teknik ini secara Hands-On. Sebanyak 12 guru yang mewakili 12 SMA di kota Mataram ikut serta dalam pelatihan ini.  Pengenalan teknik PCR dilakukan dalam dua fase: presentasi di kelas dan demonstrasi di laboratorium. Presentasi dikelas menjelaskan dasar materi genetik dan teknik PCR dan dilanjutkan dengan demonstrasi pembuatan PCR mix, menjalankan thermal cycler dan dilanjutkan pembacaan hasil PCR di gel agarose. Peningkatan nilai pretest dan post test terjadi pada seluruh peserta dengan rerata pretest 44,4% (nilai terendah 10% dan tertinggi 70%) meningkat menjadi 83.33% (nilai terendah 70 dan nilai tertinggi 100%). Melihat antusiasme partisipan dalam kegiatan ini maka perlu dilakukan kegiatan serupa pada guru SMA yang belum terpapar dengan kegiatan ini

scholarly journals Design and development of polymerase chain reaction thermal cycler using proportional-integral temperature controller

2018 ◽  
Vol 14 (2) ◽  
pp. 213-218
Author(s):  
Chong Kim Soon ◽  
Nawoor Anusha Devi ◽  
Kok Beng Gan ◽  
Sue-Mian Then
Keyword(s):  
Polymerase Chain Reaction ◽  
Low Cost ◽  
Maximum Temperature ◽  
Gradient System ◽  
Chain Reaction ◽  
Temperature Controller ◽  
Proportional Integral ◽  
Integral Temperature ◽  
Polymerase Chain ◽  
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

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

1992 ◽  
Vol 40 (3) ◽  
pp. 333-341 ◽  
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.

An intensive hands-on course designed to teach molecular biology techniques to physiology graduate students

2002 ◽  
Vol 26 (1) ◽  
pp. 42-49 ◽  
Author(s):  
Andrea D. Weston ◽  
Sasha Stasko ◽  
Gerald M. Kidder
Keyword(s):  
Polymerase Chain Reaction ◽  
Graduate Students ◽  
Molecular Biology ◽  
Historical Perspective ◽  
Rna Expression ◽  
Chain Reaction ◽  
Hands On ◽  
Similar Information ◽  
Polymerase Chain ◽  
Intensive Course

To address a growing need to make research trainees in physiology comfortable with the tools of molecular biology, we have developed a laboratory-intensive course designed for graduate students. This course is offered to a small group of students over a three-week period and is organized such that comprehensive background lectures are coupled with extensive hands-on experience. The course is divided into seven modules, each organized by a faculty member who has particular expertise in the area covered by that module. The modules focus on basic methods such as cDNA subcloning, sequencing, gene transfer, polymerase chain reaction, and protein and RNA expression analysis. Each module begins with a lecture that introduces the technique in detail by providing a historical perspective, describing both the uses and limitations of that technique, and comparing the method with others that yield similar information. Most of the lectures are followed by a laboratory session during which students follow protocols that were carefully designed to avoid pitfalls. Throughout these laboratory sessions, students are given an appreciation of the importance of proper technique and accuracy. Communication among the students, faculty, and the assistant coordinator is focused on when and why each procedure would be used, the importance of each step in the procedure, and approaches to troubleshooting. The course ends with an exam that is designed to test the students’ general understanding of each module and their ability to apply the various techniques to physiological questions.

scholarly journals An Innovative Rapid Thermal Cycling Device for Polymerase Chain Reaction

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.

Portable plastic syringe as a self-actuated pump for long-distance uniform delivery of liquid inside a microchannel and its application for flow-through polymerase chain reaction on chip

RSC Advances ◽  
2015 ◽  
Vol 5 (16) ◽  
pp. 12071-12077 ◽  
Author(s):  
Wenming Wu ◽  
Kieu The Loan Trinh ◽  
Yu Zhang ◽  
Nae Yoon Lee
Keyword(s):  
Polymerase Chain Reaction ◽  
Flow Rate ◽  
Chain Reaction ◽  
Long Distance ◽  
Chip Flow ◽  
Hands On ◽  
Plastic Syringe ◽  
Flow Through ◽  
Polymerase Chain ◽  
On Chip

A strategy for realizing self-actuated pumping with uniform flow rate over a long distance is introduced using hands-on operation of disposable syringe, and was applied for on-chip flow-through PCR inside a serpentine PMMA microchannel.

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

2002 ◽  
Vol 85 (5) ◽  
pp. 1025-1028 ◽  
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.

Failure Prediction of Polymerase Chain Reaction Thermal Cycler

2017 ◽  
Author(s):  
Mi-So Lee ◽  
Chan-Young Park ◽  
Yu-Seop Kim ◽  
Hye-Jeong Song ◽  
Jong-Dae Kim
Keyword(s):  
Polymerase Chain Reaction ◽  
Failure Prediction ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Thermal Cycler

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

2016 ◽  
Vol 6 (11) ◽  
pp. 328 ◽  
Author(s):  
Chan-Young Park ◽  
Jae-Hyeon Cho ◽  
Yu-Seop Kim ◽  
Hye-Jeong Song ◽  
Jong-Dae Kim
Keyword(s):  
Polymerase Chain Reaction ◽  
Low Cost ◽  
Temperature Logger ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Thermal Cycler
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