PaperClip DNA Assembly: Reduce, Reuse, Recycle
| Autor: | Uma Jayachandran, Alistair Elfick, Annegret Honsbein, Maryia Trubitsyna, Christopher E. French |
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| Rok vydání: | 2020 |
| Předmět: |
Cloning
0303 health sciences Oligonucleotide Computer science 030303 biophysics Pcr cloning Transfection Computational biology Molecular cloning Reuse Yeast 03 medical and health sciences Synthetic biology Transformation (genetics) chemistry.chemical_compound Plasmid chemistry Agarose gel electrophoresis Dna assembly Ligation DNA 030304 developmental biology |
| Zdroj: | Methods in Molecular Biology ISBN: 9781071609071 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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