Biotin-coupled mirna capture

WebDec 18, 2024 · Biotin-coupled miRNA capture. The 3′ end biotinylated miRNA mimics or control RNA (Ribio, Guangzhou, China) were transfected into ccRCC cells for 48 h before harvest. Then biotin-coupled RNA complex was pulled down by incubating the cell lysates with streptavidin-coated magnetic beads (Life Technologies). The abundance of … WebJan 15, 2024 · Biotin-coupled circRNA and miRNA capture. Biotin-coupled miRNA and circRNA pull-down assays were performed as described previously [11, 12]. Briefly, 3′ end biotinylated circRNA-ACAP2 (digoxin-5′-TCACACAGGCAGCTTTGGAAGAAG-3′-Digoxin; RiboBio, Guangzhou, China) was transfected into SW480 cells at a final concentration of …

Silencing of circular RNA‑ZYG11B exerts a neuroprotective effect ...

WebNov 17, 2024 · In addition, a biotin-coupled miRNA capture assay was performed using a biotin-coupled miR-185-3p probe, followed by qRT-PCR. It was observed a fourfold enrichment of circ_0008494 in the miR … WebMiR-4306 response elements were predicted by bioinformatics algorithm and identified using dual-luciferase reporter, RNA immunoprecipitation, and biotin-coupled miRNA capture assays. Results CircPDE4D was markedly downregulated in OA cartilages and interleukin (IL)-1β-stressed human normal chondrocytes (HNC). c \u0026 h the sweep https://kenkesslermd.com

Biotin-based Pulldown Assay to Validate mRNA Targets of

WebNov 9, 2024 · On the contrary, some studies argued that biotin moiety when attached to the 3′ end of the miRNA impede its association with AGO2. 41 Hence, biotin-labelled miRNA was not co-IPed with AGO2. 28 The negative result could be due to low transfection efficiency or post-translational modification of AGO2 that can hamper miRNA-AGO2 … WebJun 19, 2024 · Biotin-coupled miRNA capture. The biotin-coupled miRNA pull-down assay was performed as previously described 11. Briefly, 3′-end biotinylated miR-587 mimics (Ribio, Guangzhou, China) were ... WebJun 21, 2024 · In addition, luciferase reporter, biotin‑coupled miRNA capture and RNA immunoprecipitation assays revealed that circZYG11B could regulate RGC function through circZYG11B/microRNA‑620/PTEN signaling. Clinically, RT‑qPCR assays demonstrated that circZYG11B expression was markedly increased in the aqueous humor of patients with … c\u0026h tree surgery ltd

circCHST15 is a novel prognostic biomarker that promotes clear …

Category:mRNAs biotinylated within the 5′ cap and protected against …

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Biotin-coupled mirna capture

The TWIST1-centered competing endogenous RNA …

WebFeb 27, 2013 · Biotin-coupled miRNA capture Biotin-labelled miRNAs were transfected into HEK293 cells at a final concentration of 20 nM. Forty-eight hours after transfection, cells were collected and treated as ... WebAug 7, 2024 · The capture probe, 35 to 50 base-long and coupled to biotin on its 3′ end, binds the 5′ half of the miRNA. The detection probe is composed of a 3′ end complementary to the miRNA and a chain of fluorescently labeled RNA sequences, forming a fluorescent barcode associated with the target miRNA.

Biotin-coupled mirna capture

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WebFeb 19, 2024 · Provided herein are methods of identifying the spatial location of a nucleic acid in a biological sample. In some embodiments, the methods employ a template switching oligonucleotide. In some embodiments, the methods extend the capture probe to create a complementary DNA (cDNA) molecule of a capture analyte and produce second strand … WebJul 19, 2024 · Bioinformatic analysis was utilized to predict potential miRNA target sites, and biotin-coupled miRNA capture, biotin-coupled probe pull-down assay, and RNA …

WebMay 26, 2024 · We conducted biotin-coupled miRNA capture and FISH to confirm the sponge effect of Cdr1as. Biotin-coupled miR-1270 analogously captured more Cdr1as than biotin-coupled NC. This phenomenon indicates that miR-1270 could bind to Cdr1as (Fig. 3G). PCR experiments were conducted to find out the relative abundance of miR-1270 in … WebJan 10, 2024 · Biotin‐coupled miRNA capture The 50 μ m biotinylated miRNA mimic (GenePharma, Shanghai, China, sequence GCCCTTCATGCTGCCCAG) was …

WebIn the biotin-coupled miRNA capture assay, the direct interaction between HIPK3 and miR-194 was verified. While upregulating HIPK3 promoted NSCLC cell proliferation, co-transfection with both HIPK3 and miR-194 abrogated the tumor-promoting effects of HIPK3. Altogether, these data corroborated that HIPK3 and miR-149 exert opposing roles in … WebAug 17, 2024 · To validate the interactions between circCDYL and its targets, we performed RNA fluorescent in situ hybridization, biotin-coupled miRNA capture assay and biotin-coupled probe pull-down assay. Tight …

WebNov 23, 2024 · Biotin-coupled miRNA capture, fluorescence in situ hybridization and luciferase reporter assays were conducted to confirm the relationship between circMMP9 and miR-124. In this study, we screened differentially expressed circRNAs and identified circMMP9 in GBM. We found that circMMP9 acted as an oncogene, was upregulated in …

WebJun 25, 2024 · Biotin-coupled miRNA capture. The biotin-coupled miRNA pull-down assay was performed using previously reported method 8. Briefly, the 3′-end biotinylated … c\u0026h taxi charleston wvWebDec 20, 2024 · Biotin-coupled miRNA capture. For biotin-coupled capture, approximately 5 × 10 6 BCa cells (T24 and 253 J-BV) were transfected with biotin-miR135a-5p or negative control (NC) by … eassist dental billing test answersWebJul 13, 2024 · 2.13 Biotin-coupled miRNA capture. The biotin-coupled miRNA pull-down assay was carried out as previously described. 17, 18 Briefly, the 3′-end biotinylated miR-RNA mimic or control biotin-RNA (RiboBio) was transfected into SPCA1 cells at a final concentration of 20 nmol/L for 1 day. The biotin-coupled RNA complex was pulled … eassist eobWebInvitrogen streptavidin-coupled Dynabeads are a robust and versatile tool that can be used to capture specific RNA or DNA sequences and then pull them directly out of solution. … eassist insurance verificationWebDec 12, 2024 · Streptavidin coupled to a solid matrix of beads allows the specific capture of biotinylated RNA easily. To demonstrate the suitability of biotinylated cap analogues for the study of protein–RNA interactions we established a combined protocol including ultraviolet (UV) cross-linking, and affinity purification of biotinylated-capped RNA. c\u0026h truck parts westmoreland tnWebJan 16, 2024 · RNA immunoprecipitation (RIP) and biotin-coupled miRNA capture. g for 20 min. Western blot analysis. The total protein of 95D cells were exacted using protein extraction reagent (Thermo Scientific ... eassist insuranceWebFeb 27, 2013 · Biotin-coupled miRNA capture Biotin-labelled miRNAs were transfected into HEK293 cells at a final concentration of 20 nM. Forty-eight hours after transfection, cells were collected and treated as ... c \u0026 h tools peoria il