Ethidium Bromide Etbr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "A PCR-free rapid protocol for one-pot construction of highly diverse genetic libraries"
Article Title: A PCR-free rapid protocol for one-pot construction of highly diverse genetic libraries
Journal: PLOS ONE
Figure Legend Snippet: Library preparation results. (A) Single-stranded DNA products of Step 1 and 2 visualized on a 15% TBE-Urea gel stained with SYBR Green. Yellow arrows mark the three-way ligation products of Step 1, and the green arrow marks the presumed complete ligation product after combining all three groups. (B) Double-stranded DNA products from Step 3 visualized on a 1% agarose gel stained with ethidium bromide. Lanes 1 and 2 used DNA template of Ligation 2 (Step 2) produced in the presence and absence of ligase, respectively. Lane 3 (PC) contains gel-purified full-length HARPin double-stranded DNA (693 bp) amplified by PCR as a positive control. (C) mRNA product from Step 4 visualized on a denaturing 1.5% agarose gel stained with EtBr (1.5% agarose, 1% bleach). Lanes 1–3 contained the corresponding transcription product from panel B. The full-length mRNA contains 669 nt. The gels are representative of three independent experiments.
Techniques Used: Staining, SYBR Green Assay, Ligation, Agarose Gel Electrophoresis, Produced, Purification, Amplification, Polymerase Chain Reaction, Positive Control
2) Product Images from "Three-Layered Complex Interactions among Capsidless (+)ssRNA Yadokariviruses, dsRNA Viruses, and a Fungus"
Article Title: Three-Layered Complex Interactions among Capsidless (+)ssRNA Yadokariviruses, dsRNA Viruses, and a Fungus
Figure Legend Snippet: Identification of the partner of YkV3 and YkV4. (A) Confirmation of single virus infection in R. necatrix W97 transfected with RnMBV3, RnMTV1s, or RnFGV3 by RT-PCR. Strain W97 is a virus-free control. The field-collected fungal strains Rn454 and Rn95-16 as a source of the viruses were analyzed in parallel. Rntub2 ( R. necatrix β -tubulin gene) mRNA served as an internal control. (B) Experimental flow to identify dsRNA virus partners that support the replication of yadokariviruses. T-YkV n represents R. necatrix W97 transformed with an infectious cDNA clone of each yadokarivirus (where n refers to the numbers attached to the names of yadokariviruses). The transformants (T-YkV n ) were cocultured for approximately 10 days with each of the transfectants harboring a single candidate dsRNA virus. Antibiotic resistant T-YkV n sides and antibiotic susceptible viral transfectant sides were each subcultured to examine for yadokarivirus replication and horizontal virus transfer. When yadokarivirus replication was supported, coculture allowed for mutually lateral virus transfer in both directions via hyphal fusions. If not, only dsRNA viruses could be transferred from the transfectant side to the transformant side. Yadokarivirus replicative form dsRNA was detected in R. necatrix subcolonies before and after the coculture between T-YkV n and the dsRNA virus transfectants. (C to E) The three coculture pairs were tested: the YkV3 cDNA transformant (T-YkV3) and the RnMBV3 transfectant (C); the YkV4a cDNA transformant (T-YkV4a) and the RnMTV1a transfectant (D); the YkV4b cDNA transformant (T-YkV4b) and the RnMTV1b transfectant (E). The top two panels show total or yadokarivirus dsRNA detected by ethidium bromide or Northern hybridization, respectively. The black triangles indicate dsRNA of RnMBV3 or RnMTV1s. The red triangles indicate dsRNA of YkV3 or YkV4s. The asterisks indicate defective dsRNA of RnMTV1. The values below the total dsRNA panel indicate the abundance ratio of the yadokarivirus dsRNA to the partner viral dsRNA measured by ImageJ in which the partner dsRNA intensity was expressed as 1. The middle two panels show PCR-based detection of the transgene of yadokariviruses cDNA and an internal control, respectively. A host β-tubulin gene ( Rntub2 DNA) was detected as an internal control. The bottom panel shows the electrophoretic profile of a portion of total nucleic acid (1 μg), the batch of which was used for dsRNA extraction (top two panels) and host genotyping (middle panels).
Techniques Used: Infection, Transfection, Reverse Transcription Polymerase Chain Reaction, Transformation Assay, Northern Blot, Hybridization, Polymerase Chain Reaction, Genotyping Assay
3) Product Images from "Targeted introduction of heritable point mutations into the plant mitochondrial genome"
Article Title: Targeted introduction of heritable point mutations into the plant mitochondrial genome
Journal: Nature plants
Figure Legend Snippet: Workflow for the isolation of tobacco lines with TALEN-induced point mutations in the mitochondrial nad9 gene. ( a ) Schematic overview of the experimental procedures. Transgenic plants with confirmed TALEN activity (cf. Extended Data Fig. 1 ) were raised from seeds, leaves were harvested, cut into pieces and placed onto shoot induction medium. Leaves from regenerated shoots were genotyped for mutations in nad9 (‘Discovery of mutation’). If no mutation was detected, a new regeneration round was initiated. When a point mutation was found, an additional regeneration round was conducted to promote genome segregation and facilitate isolation of homochondriomic lines (‘Purification of mutation’). Homochondriomic mutant shoots were rooted and transferred to the greenhouse for seed production. In some experiments, the mutagens (M) ethidium bromide or N-nitroso-N-ethylurea were added to the shoot induction medium or applied during seed imbibition. See text, Methods and Supplementary Methods for details. ( b ) Exemplary sequencing chromatograms showing the successful isolation of a homochondriomic nad9 mutant carrying a Ser-to-Gly exchange in amino acid position 91 of the Nad9 protein (corresponding to an A-to-G substitution in nucleotide position 271 of the nad9 reading frame). The mutated position is indicated by arrowheads in the sequencing chromatograms of the S91G-1 mutant line and the wild type. The sequencing primer was oJF271 ( Supplementary Table 2 ).
Techniques Used: Isolation, Transgenic Assay, Activity Assay, Mutagenesis, Purification, Sequencing
4) Product Images from "P2RX7 inhibition reduces breast cancer induced osteolytic lesions - implications for bone metastasis"
Article Title: P2RX7 inhibition reduces breast cancer induced osteolytic lesions - implications for bone metastasis
Figure Legend Snippet: A) mRNA expression for P2RX7 in the different molecular subtypes of breast cancer. Data analysed from the TCGA and SCAN-B datasets (n=4669). B) mRNA expression of P2RX7 in triple-negative breast cancer (TNBC, n=317) compared to non-TNBC (n=4119). C) Kaplan-Meier plot for distant-metastasis free survival for patients with low (tercile 1) and high (tercile 3) P2RX7 expression. D) P2RX7 agonist BzATP (100μM) mediated calcium response (green) in E0771 murine breast cancer cells, which is inhibited in presence of a specific antagonist A740003 (10μM, red). Peak intensity and the area-under-curve (AUC) for the calcium response in (D) are presented in (E) and (F) respectively. G) P2RX7 mediated pore formation with higher concentrations of BzATP (300μM) assessed by ethidium bromide uptake in E0771 cells (red), which is inhibited by A740003 (green). P2RX7 over-expressing HEK-293 cells are used as a positive control (blue). (H) Analysis of the AUC for these pore-formation responses. I) Effect of P2RX7 antagonist A740003 on E0771 cell proliferation assessed by WST-1 at 72h. J) The effect of A740003 (10μM) on E0771 migration using scratch assay. All in vitro data is presented relative to the untreated controls and are from 3 separate biological repeats. The data has been presented as mean ± SD and analysed using one-way ANOVA. *P
Techniques Used: Expressing, Positive Control, Migration, Wound Healing Assay, In Vitro
5) Product Images from "Cellular Growth Arrest and Efflux Pumps Are Associated With Antibiotic Persisters in Streptococcus pyogenes Induced in Biofilm-Like Environments"
Article Title: Cellular Growth Arrest and Efflux Pumps Are Associated With Antibiotic Persisters in Streptococcus pyogenes Induced in Biofilm-Like Environments
Journal: Frontiers in Microbiology
Figure Legend Snippet: Streptococcus pyogenes cells in biofilm-like environments show persistence to ethidium bromide and to a number of non-β-lactam antimicrobials. (A) Persisters recovered from biofilm-like environments at concentrations of 4 μg/mL ethidium bromide (EtBr; MIC = 0.06 μg/mL), 4 μg/mL erythromycin (Ery; MIC = 0.12 μg/mL), 4 μg/mL azithromycin (Azi; MIC = 0.12 μg/mL), 1 μg/mL clindamycin (Cli; MIC = 0.01 μg/mL), 16 μg/mL chloramphenicol (Chl; MIC = 1 μg/mL), or 16 μg/mL tetracycline (Tet; MIC = 0.12 μg/mL). The average CFU/mL of the control cells (no antibiotic) was 5.5 × 10 10 and corresponded to 100%. One way ANOVA was applied using CFU values ( p