Journal: bioRxiv

Article Title: A mRNA-LNP vaccine against Dengue Virus elicits robust, serotype-specific immunity

doi: 10.1101/2021.01.05.425517

Figure Lengend Snippet: A) Schematic of the DENV genome and engineered mRNA construct. An mRNA encoding for the prM and ENV viral proteins was engineered with N-terminal signal peptide sequence, 5’ and 3’ untranslated regions (UTR) flanking the coding sequence, a 3’ poly-A tail, and a 5’ cap-1 structure. In vitro synthesized mRNA is encapsulated in a lipid nanoparticle for use in in vitro and in vivo experiments. B) 293T cells were transfected with the in vitro transcribed mRNA encoding for the wild-type sequence (WT), or a mutant version with amino acid substitutions in the fusion-loop epitope (ΔFL). Lysate was analyzed by western blot with the domain III specific 1A1D-2 monoclonal antibody and the fusion-loop specific 4G2 monoclonal antibody. C) Supernatant from transfected cells was purified and concentrated through ultracentrifugation and analyzed for VLPs by western blots with the 1A1D-2 monoclonal antibody or anti-GAPDH. Unpurified cell lysate from WT mRNA transfected cells is included as a control. Shown are representative blots. D) Electron microscopy image of VLPs from purified supernatant of transfected 293T cells showing homogenous shape and size of approximately 30nm.

Article Snippet: Membranes were blotted with envelope domain III specific 1A1D-2 (1:600) monoclonal antibody (CDC Arbovirus Reference Collection) or envelope fusion-loop specific 4G2 (3.33 mg/ml) (BEI Cat# NR-50327, Novus Biologicals Cat# NBP2-52709FR).

Techniques: Construct, Sequencing, In Vitro, Synthesized, In Vivo, Transfection, Mutagenesis, Western Blot, Purification, Control, Electron Microscopy

Journal: Genes

Article Title: Improved DNA Extraction and Illumina Sequencing of DNA Recovered from Aged Rootless Hair Shafts Found in Relics Associated with the Romanov Family

doi: 10.3390/genes13020202

Figure Lengend Snippet: Description of the hair shaft samples that were used in this study and purification methods that were used during DNA extraction.

Article Snippet: The MinElute columns were, therefore, replaced first with the Ultraclean Production (UCP) MinElute columns (cat # 56204, Qiagen).

Techniques: Purification, Extraction

Journal: Genes

Article Title: Improved DNA Extraction and Illumina Sequencing of DNA Recovered from Aged Rootless Hair Shafts Found in Relics Associated with the Romanov Family

doi: 10.3390/genes13020202

Figure Lengend Snippet: Bioanalyzer electropherograms of reagent blank libraries when DNA purification was performed using ( A ) MinElute columns, or ( B ) magnetic beads from G-Biosciences.

Article Snippet: The MinElute columns were, therefore, replaced first with the Ultraclean Production (UCP) MinElute columns (cat # 56204, Qiagen).

Techniques: DNA Purification, Magnetic Beads

Journal: mBio

Article Title: Dynamics of Mycobacterium tuberculosis Ag85B Revealed by a Sensitive Enzyme-Linked Immunosorbent Assay

doi: 10.1128/mBio.00611-19

Figure Lengend Snippet: Recognition of Ag85A, Ag85B, and Ag85C by monoclonal antibodies 710, 711, and 712. (A to C) Wells of 96-well plates were coated with Ag85B (A), Ag85A (B), or Ag85C (C), each at 0.5 µg/ml. mAb 710, 711, or 712 was added at the indicated concentrations, and binding was detected after washing, using HRP-conjugated goat anti-mouse IgG and the TMB substrate. Data shown in panels A to C are representative of results from three independent experiments with one technical replicate per experiment per condition. (D to F) Individual wells were coated with a dilution series of Ag85B (D), Ag85A (E), or Ag85C (F), starting at 20 µg/ml. mAb 710, 711, or 712 was added at a concentration of 1 µg/ml, and binding was detected after washing, using HRP-conjugated goat anti-mouse IgG and the TMB substrate. Data in panels D to F are representative of results from two independent experiments with one technical replicate per experiment per condition.

Article Snippet: Ag85A (catalog number NR-14871; BEI Resources), Ag85B (our purified recombinant), or Ag85C (catalog number NR-14858; BEI Resources) was used to coat wells at 0.5 μg/well in phosphate-buffered saline (PBS) and incubated overnight at 4°C.

Techniques: Bioprocessing, Binding Assay, Concentration Assay