PaperClip DNA assembly: Reduce, reuse, recycle

Maryia Trubitsyna, Annegret Honsbein, Uma Jayachandran, Alistair Elfick, Christopher E. French

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Creating DNA constructs is a basic and fundamental step in molecular and synthetic biology. While prices for gene synthesis are decreasing, it is still more economical in most cases to assemble constructs from a library of components (Parts). Many methods for DNA assembly are available, but most require either a fixed and inflexible format for the construct, with all Parts first being cloned in specific donor plasmids, or remaking Parts with new homology ends for each specific assembly reaction, requiring large numbers of single-use oligonucleotides. PaperClip assembly allows Parts stored in any format (linear PCR products or synthetic DNA, or cloned in any plasmid) to be used in totally flexible assembly reactions; up to 11 parts can be assembled in a single reaction, in any order, to give a linear or circular construct, and the oligonucleotides required in the assembly process can be reused in any subsequent assembly. In addition to constructing plasmids for bacterial transformation, PaperClip is also well suited to generate linear products for direct transfection of yeast, mammalian, or cyanobacterial cell lines. Thus, PaperClip offers a simple, flexible, and economical route to multipart assembly of constructs for a wide variety of purposes.
Original languageEnglish
Title of host publicationDNA Cloning and Assembly
PublisherSpringer
Chapter10
Pages161-177
Volume2205
ISBN (Electronic)978-1-0716-0908-8
ISBN (Print)978-1-0716-0907-1
DOIs
Publication statusPublished - 19 Aug 2020

Publication series

NameDNA Cloning and Assembly
Volume2205
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • synthetic biology
  • molecular cloning
  • DNA assembly
  • single-pot assembly
  • oligonucleotide annealing
  • ligation
  • PAGE
  • PCR
  • agarose gel electrophoresis
  • CPEC
  • selection

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