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Related Concept Videos

PCR01:32

PCR

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DNA Isolation01:24

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...
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Next-generation Sequencing

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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

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Sanger Sequencing

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Related Experiment Video

Updated: May 12, 2026

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
07:35

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems

Published on: June 14, 2021

A simple, universal, efficient PCR-based gene synthesis method: sequential OE-PCR gene synthesis.

Pingping Zhang1, Yingying Ding, Wenting Liao

  • 1Department of Medical Microbiology, Second Military Medical University, 800 Xiangyin Road, Shanghai 200433, China.

Gene
|April 20, 2013
PubMed
Summary

This study introduces a straightforward gene synthesis method using sequential overlap extension polymerase chain reactions (OE-PCR). The technique efficiently produces full-length genes, overcoming common obstacles like mispriming and secondary structures.

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Last Updated: May 12, 2026

Rapid, Enzymatic Methods for Amplification of Minimal, Linear Templates for Protein Prototyping using Cell-Free Systems
07:35

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Published on: June 14, 2021

Multi-target Parallel Processing Approach for Gene-to-structure Determination of the Influenza Polymerase PB2 Subunit
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Multi-target Parallel Processing Approach for Gene-to-structure Determination of the Influenza Polymerase PB2 Subunit

Published on: June 28, 2013

Area of Science:

  • Molecular Biology
  • Synthetic Biology
  • Biotechnology

Background:

  • Gene synthesis is crucial for molecular biology research and biotechnology.
  • Existing methods can be complex, time-consuming, or require specialized expertise.
  • Challenges include mispriming and secondary structures that impede successful synthesis.

Purpose of the Study:

  • To present a simple, universal, and efficient gene synthesis method.
  • To provide an alternative tool for individual gene synthesis.
  • To address and overcome common obstacles in gene synthesis.

Main Methods:

  • Utilizing sequential overlap extension polymerase chain reactions (OE-PCR).
  • Designing paired complementary 54-mer oligonucleotides with 18 bp overlaps.
  • Employing sequential OE-PCR for full-length gene synthesis, followed by cloning, sequencing, and resynthesis with correct templates.

Main Results:

  • The method successfully synthesizes full-length genes.
  • Mispriming and secondary structures were identified as key obstacles and were effectively resolved.
  • The process allows for high error tolerance in polymerase, oligonucleotides, and PCR conditions.

Conclusions:

  • This OE-PCR based method offers a reliable and efficient approach to gene synthesis.
  • It provides a valuable alternative for researchers without specialized gene synthesis experience.
  • The method guarantees successful gene synthesis in most cases, enhancing accessibility.