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PCR

PCR

PCR - Polymerase Chain Reaction

PCR - Polymerase Chain Reaction

Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

DNA Isolation

DNA Isolation

DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...

RACE - Rapid Amplification of cDNA Ends

RACE - Rapid Amplification of cDNA Ends

Rapid Amplification of cDNA Ends, or RACE, is one of the most effective methods to obtain a full-length cDNA from an mRNA sequence between a known internal region to the unknown sequence at the 5’ or 3’ end. The unknown region is cloned in the cDNA by a gene-specific primer that binds the known end, and a hybrid primer that attaches a predefined anchor sequence to the unknown end of the cDNA. The sequence in between is amplified by PCR with an anchor primer and a gene-specific...

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Related Experiment Video

Updated: May 5, 2026

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies

Published on: May 22, 2012

The polymerase chain reaction (PCR): general methods.

Daniel L E Waters¹, Frances M Shapter

¹Southern Cross Plant Science, Southern Cross University, Lismore, NSW, Australia.

Methods in Molecular Biology (Clifton, N.J.)

|November 19, 2013

Summary

This summary is machine-generated.

The polymerase chain reaction (PCR) amplifies minute DNA amounts into large quantities, forming the basis for molecular biology techniques. This method uses cellular machinery in vitro for predictable DNA amplification based on user input.

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Area of Science:

Molecular Biology
Biotechnology

Background:

The polymerase chain reaction (PCR) is a fundamental technique in molecular biology.
PCR enables the amplification of specific DNA sequences from minute quantities.

Purpose of the Study:

To introduce the principles of the polymerase chain reaction (PCR).
To discuss practical considerations for PCR implementation, including target sequence definition, optimization, and applications.

Main Methods:

The chapter explains how PCR utilizes in vitro components of cellular mitotic division machinery.
It highlights the predictable response of these components to user-defined inputs.

Main Results:

PCR effectively converts very low quantities of DNA into very high quantities.
The principles discussed enable a wide range of specialized molecular biology techniques.

Conclusions:

Understanding PCR principles is crucial for its effective application in molecular biology.
Practical considerations are essential for successful DNA amplification and downstream applications.