title: One primer to rule them all: universal primer that adds BBa_B0034 ribosomal binding site to any coding standard 10 BioBrick.
abstract: Here, we present a universal, simple, efficient, and reliable way to add small BioBrick parts to any BioBrick via PCR that is compatible with BioBrick assembly standard 10. As a proof of principle, we have designed a universal primer, rbs_B0034, that contains a ribosomal binding site (RBS; BBa_B0034) and that can be used in PCR to amplify any coding BioBrick that starts with ATG. We performed test PCRs with rbs_B0034 on 31 different targets and found it to be 93.6% efficient. Moreover, when supplemented with a complementary primer, addition of RBS can be accomplished via whole plasmid site-directed mutagenesis, thus reducing the time required for further assembly of composite parts. The described method brings simplicity to the addition of small parts, such as regulatory elements to existing BioBricks. The final product of the PCR assembly is indistinguishable from the standard or 3A BioBrick assembly.
authors.affiliation: Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University , Atlanta, Georgia 30332, United States.
authors.forename: Anton V
authors.initials: AV
authors.lastname: Bryksin
authors.forename: Haylee N
authors.initials: HN
authors.lastname: Bachman
authors.forename: Spencer W
authors.initials: SW
authors.lastname: Cooper
authors.forename: Tilak
authors.initials: T
authors.lastname: Balavijayan
authors.forename: Rachael M
authors.initials: RM
authors.lastname: Blackstone
authors.forename: Haoli
authors.initials: H
authors.lastname: Du
authors.forename: Jackson P
authors.initials: JP
authors.lastname: Jenkins
authors.forename: Casey L
authors.initials: CL
authors.lastname: Haynes
authors.forename: Jessica L
authors.initials: JL
authors.lastname: Siemer
authors.forename: Vincent F
authors.initials: VF
authors.lastname: Fiore
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Barker
mesh_terms: D001665:Binding Sites / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics*; D008967:Molecular Biology; D016297:Mutagenesis, Site-Directed; D010957:Plasmids / Q000235:genetics; D012270:Ribosomes / Q000235:genetics* / Q000378:metabolism
chemical_list: D017931:DNA Primers