Megaprimer-based PCR to synthesize fusion genes for cloning

Tatiana Q. Aguiar*, Carla Oliveira, Lucília Domingues

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Citation (Scopus)

Abstract

Megaprimer-based polymerase chain reaction (PCR) strategies allow the versatile and fast assembly and amplification of a myriad of tailor-made or random DNA sequences readily available for conventional or restriction-free (RF) cloning. In this chapter, we present a megaprimer-based PCR protocol that enables the expeditious construction of customized fusion genes ready for cloning into commercial expression plasmids. With the expanding use of protein tag technology in the most diverse application fields, this protocol remains a versatile and affordable solution for the synthesis and fusion of peptide tags/domains of interest.
Original languageEnglish
Title of host publicationPCR
Subtitle of host publicationmethods and protocols
EditorsLucília Domingues
Place of PublicationNew York
PublisherHumana Press Inc.
Pages193-207
Number of pages15
Edition2
ISBN (Electronic)9781071633588
ISBN (Print)9781071633571, 9781071633601
DOIs
Publication statusPublished - 23 Aug 2023

Publication series

NameMethods in Molecular Biology
Volume2967
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Megaprimers
  • PCR
  • Fusion genes
  • Molecular cloning
  • Restriction enzymes
  • Expression plasmids

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