Restriction enzymes and β agarase

Recombinant DNA methods were in accordance with established procedures and used commercially available reagents: Phusion High-Fidelity DNA Polymerase (Thermo Fisher, Waltham, MA); restriction enzymes and β-agarase (New England BioLabs, Beverly, MA); T4 DNA ligase, CIP, and T4 PNK (Roche, Nutley, NJ); GTG low melting temperature agarose for in-gel cloning, (Lonza, Walkersville, MD); oligonucleotides and gBlocks® (Integrated DNA Technologies, Coralville, IA); Assemblies involving polymerase chain reaction (PCR) amplification were sequenced through the inserts and junctions to verify the desired construct. Cloning was carried out in XL1-Blue cells. Full details of cloning, oligonucleotides, maps, and sequences of the resulting constructs are available on request.

Recombinant DNA methods were according to established procedures and employed commercially available reagents; Phusion High-Fidelity DNA Polymerase (Thermo Fisher Scientific, Waltham, MA, USA) was used for PCR amplification unless otherwise noted; restriction enzymes and β-agarase (New England BioLabs, Beverly, MA, USA); T4 DNA ligase, CIP and T4 PNK (Roche, Nutley, NJ, USA), GTG low-melting temperature agarose for in agarose cloning (Lonza, Walkersville, MD, USA); oligonucleotides (Integrated DNA Technologies, Coralville, IA, USA); synthetic human codon optimized sdAb and VP40 genes (Genscript, Piscataway, NJ, USA). Assemblies involving cloning and PCR amplification were sequenced through the inserts and junctions to verify the desired construct. Cloning was typically in XL1-Blue unless otherwise stated. Full details of cloning, oligonucleotides, maps, and sequences of resulting constructs are available on request. Parental sdAb genes employed in this work were anti-MARV NP sdAb A, B, C, and D with GenBank accession numbers MF780583, MF780584, MF780585, and MF780586, respectively; anti-EBOV NP sdAb E and G with GenBank accession numbers MF780602 and MF780604, respectively.