PCR Archives - #1 Microbiology Resource Hub

REAL-TIME PCR COMPONENTS

DNA polymerasePCR performance is often related to the thermostable DNA polymerase, so enzyme selection is critical to success. One of the main factors affecting PCR specificity is the fact that Taq DNA polymerase has residual activity at low temperatures. Primers can anneal nonspecifically to DNA during reaction setup, allowing the polymerase to synthesize nonspecific product. […]

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Molecular Microbiology, Techniques in Microbiology Labdeoxyadenosine triphosphate (dATP), deoxycytidine triphosphate (dCTP), deoxyguanosine triphosphate (dGTP), Deoxynucelotide triphosphates, deoxythymidine triphosphate (dTTP), DNA polymerase, dNTPs, PCR, REAL-TIME PCR COMPONENTS, Reverse transcriptase, Taq DNA polymerase

STEPS INVOLVED IN PERFORMING REAL-TIME PCR

DrChika

Real-time PCR is a variation of the standard PCR technique that is commonly used to quantify DNA or RNA in a sample. Using sequence-specific primers, the number of copies of a particular DNA or RNA sequence can be determined. By measuring the amount of amplified product at each stage during the PCR cycle, quantification is

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Molecular Microbiology, Techniques in Microbiology Labannealing, denaturation, DNA polymerase, extension, PCR, PCR reaction, primers, qRT-PCR, quantitative reverse transcriptase PCR, real time PCR, RT PCR, steps involved in real time PCR

INTRODUCTION TO REAL TIME POLYMERASE CHAIN REACTION (RT PCR)

DrChika

The polymerase chain reaction (PCR) is one of the most powerful technologies in molecular biology. Using PCR, specific sequences within a DNA or complementary DNA (cDNA) template can be copied, or “amplified”, many thousand- to a million-fold using sequence-specific oligonucleotides (or primers), heat-stable DNA polymerase, and thermal cycling. In traditional (endpoint) PCR, detection and quantification

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Biotechnology, Molecular Microbiology, Techniques in Microbiology Labamplification, cDNA, cDNA template, cycle number, gene amplification, PCR, qPCR, REAL TIME POLYMERASE CHAIN REACTION, RT PCR, RT qPCR

GENOTYPIC DETECTION OF ANTIBIOTIC RESISTANT MICROBES

DrChika

The genotypic detection and characterization of antibiotic resistant genes in pathogenic bacteria is more specific in characterizing resistance genes than the phenotypic detection methods. The term genotypic is derived from genotype – which is the complete genetic makeup of an organism. Genotype describes the overall composition of the genetic information found in an organism. Thus,

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Antimicrobial Susceptibility Testing (AST) & Antibiogram, Pharmaceutical Microbiology, Techniques in Microbiology Lab. Restriction fragment length polymorphism (RFLP), DNA microarray technique, DNA-based methods, Fluorescent in situ hybridization (FISH), GENOTYPIC DETECTION OF ANTIBIOTIC RESISTANT MICROBES, ligase – chain reaction (LCR), nucleotide sequencing, PCR, PCR-single-stranded conformation polymorphism (SSCP), Random Amplified Polymorphic DNA (RAPD)

PCR TECHNIQUE

DrChika

PCR is the acronym for “polymerase chain reaction”. Polymerase chain reaction is the molecular biology technique that is used to amplify or copy specific gene sequences or nucleotides of a DNA molecule or gene. It was invented by Kary Mullis and his colleagues in the early 1980s; and PCR technique is the most widely used

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Biotechnology, Molecular Microbiology, Techniques in Microbiology LabPCR, PCR TECHNIQUE

THERMOCYCLER (PCR Machine)

DrChika

Thermocycler or thermal cycler is a piece of equipment is used for the copying or amplification of specific sequences of nucleotides through a controlled timing and temperature regulation. It is an important tool in the molecular biology laboratory. Thermocycler allows reaction mixtures in specialized tubes known as Eppendorf tubes (PCR tubes) to be heated and

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Biotechnology, Molecular Microbiology, Techniques in Microbiology LabPCR, THERMO CYCLER, THERMOCYCLER