Journal: STAR Protocols

Article Title: Rapid and efficient enhancer cloning and in vivo screening using the developing chick embryo

doi: 10.1016/j.xpro.2021.100507

Figure Lengend Snippet: Nanostring screening of putative enhancers (A) Nanotag transcripts are detected by a customized Nanostring codeset , counts above 100 (in green, above dashed line) were determined to depict in vivo enhancer activity. (B) Negative control plasmids achieved Nanostring counts less than 100. In vivo background expression is observed in the posterior extra-embryonic region. (C) Enhancers with Nanostring count >100 are re-electroporated into early chick embryos at higher concentration such that fluorescence can be observed. Multiple enhancers carrying different fluorophores can be imaged in parallel, as shown for neural crest enhancers; NC1, NC2 and Sox10E2 ( Betancur et al., 2010 ; Simoes-Costa et al., 2012 ). Figure adapted and reprinted with permission from ( Williams et al., 2019 ).

Article Snippet: We provide a number of Nanotag reporter plasmids containing the FoxD3 neural crest enhancer (NC1) ( ) as positive control (Addgene #130570 - #130573).

Techniques: In Vivo, Activity Assay, Negative Control, Expressing, Concentration Assay, Fluorescence

Journal: Journal of Clinical Microbiology

Article Title: Close Geographic Association of Human Neoehrlichiosis and Tick Populations Carrying " Candidatus Neoehrlichia mikurensis" in Eastern Switzerland

doi: 10.1128/JCM.01955-12

Figure Lengend Snippet: Analytical sensitivity of the multiplex real-time PCR test for detection of “Ca. Neoehrlichia mikurensis,” the “Ca. Neoehrlichia,” genus, and members of the Anaplasmataceae family. (A) “Ca. Neoehrlichia mikurensis”-specific probe (Neo_spec). (B) “Ca. Neoehrlichia” genus probe (Neo_genus). (C) Anaplasmataceae family probe (Ana_family). A dilution series ranging from 105 copies to 10−1 copies of the plasmid pCNM1 containing a partial 16S rRNA gene of “Ca. Neoehrlichia mikurensis” (one copy per plasmid) was used for determining the analytic sensitivity of the probes. The experiment was performed 10 times, of which one representative run is shown. All runs showed comparable results. Rn, normalized reporter dye fluorescence; H2O, negative water control.

Article Snippet: The positive-control plasmid pCNM1 containing a 400-bp segment of the 5′ end of the 16S rRNA gene ( Escherichia coli 16S rRNA gene positions 14 to 414) was constructed using in silico design and de novo synthesis and subcloning (Genscript, CA).

Techniques: Multiplex Assay, Real-time Polymerase Chain Reaction, Plasmid Preparation, Fluorescence, Control