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Related Concept Videos

PCR01:32

PCR

Overview
RACE - Rapid Amplification of cDNA Ends02:35

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 primer.
Since the...
PCR - Polymerase Chain Reaction01:32

PCR - Polymerase Chain Reaction

Overview

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Related Experiment Video

Updated: Jun 4, 2026

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)
10:00

A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

Published on: December 26, 2018

Streamlined gene assembly PCR.

Wayne M Barnes, Elaine R Frawley

    CSH Protocols
    |March 2, 2011
    PubMed
    Summary
    This summary is machine-generated.

    This study streamlines gene and plasmid construction using a one-step Polymerase Chain Reaction (PCR) assembly method. The optimized process efficiently creates artificial gene sequences without gel purification, simplifying molecular cloning workflows.

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    Last Updated: Jun 4, 2026

    A Customizable Protocol for String Assembly gRNA Cloning (STAgR)
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    A Customizable Protocol for String Assembly gRNA Cloning (STAgR)

    Published on: December 26, 2018

    Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
    08:31

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    Published on: February 5, 2021

    Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
    10:18

    Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

    Published on: February 9, 2015

    Area of Science:

    • Molecular Biology
    • Synthetic Biology
    • Biochemistry

    Background:

    • Gene and plasmid construction are fundamental in molecular biology.
    • Traditional methods often involve multiple steps and purification processes.
    • Artificial gene synthesis enables custom protein and pathway engineering.

    Purpose of the Study:

    • To develop a streamlined Polymerase Chain Reaction (PCR) based method for gene and plasmid assembly.
    • To simplify the process of creating artificial gene sequences.
    • To reduce the number of steps and eliminate the need for gel purification in gene synthesis.

    Main Methods:

    • Utilized a single-round PCR cycling protocol (20-25 cycles) for gene assembly from short DNA fragments (as small as 40 nucleotides).
    • Designed primers that anneal to each other for initial assembly without external primers.
    • Integrated direct cloning of the PCR product without intermediate gel purification.

    Main Results:

    • Achieved efficient assembly of whole genes and plasmids in a single PCR step.
    • Obtained a single or major product band detectable by agarose gel electrophoresis.
    • Demonstrated successful direct cloning of the assembled DNA fragments.

    Conclusions:

    • The streamlined PCR assembly method significantly simplifies gene and plasmid construction.
    • This optimized protocol reduces experimental time and resources.
    • The method is effective for creating artificial gene sequences for synthetic biology applications.