Biotin-coupled mirna capture
WebFeb 12, 2024 · Biotin-coupled miRNA capture assay. The biotin-labeled circHIPK3 and control probes were synthesized by GenePharma (Shanghai, China). In brief, biotin-labeled circHIPK3 and control probes were … WebJan 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) with a cocktail of proteinase inhibitors (Roche Applied Science, Switzerland). Equal amount of total protein was separated by 10% sodium …
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 … 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 …
WebInvitrogen 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. … WebMay 26, 2024 · Biotin-coupled miRNA capture. The 3′ end of biotinylated miR-556-5p and miR-1322 mimics or control RNA (Ribio, Guangzhou, China) were transfected into 1 × 10 6 HCT116 and DLD1 cells at a final concentration of 50 nM for 48 h before harvest. Then the cell pellet was incubated with 0.7 mL lysis buffer (5 mM MgCl 2. In vivo experiment
WebMay 15, 2024 · They found that miR-193a-3p functioned as a tumor suppressor in PCa cells, through bioinformatic prediction and biotin-coupled miRNA capture studies. Functional studies showed that miR-193a-3p reduced MCL1 expression, to suppress proliferation, migration, and invasion in cancer cells . WebDec 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.
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.
WebMay 31, 2024 · The biotin-coupled miRNA capture assay was performed as previously described. 17 In brief, miR-206 mimics and miRNA negative control were labeled with biotin at the 3′ end and transiently transfected into LoVo and HCT116 cells. After 24 hrs of culture, the cells were harvested and lysed in a lysis buffer, followed by incubation with ... how do you use artbreederWebJan 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 ... phoning overseas from ukWebJan 10, 2024 · Biotin‐coupled miRNA capture The 50 μ m biotinylated miRNA mimic (GenePharma, Shanghai, China, sequence GCCCTTCATGCTGCCCAG) was … phoning paris from ukWebJun 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 ... phoning over the internetWebJan 31, 2024 · Biotin-coupled probe pull down assay, Biotin-coupled miRNA capture, Fluorescence in situ hybridization and Luciferase reporter assay were conducted to confirm the relationship between the circ-ITCH and the microRNA. Results: In the present study, we found that circ-ITCH, is down-regulated in BCa tissues and cell lines. BCa patients with … phoning outside australiaWebJun 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 ... how do you use associative propertyWebNov 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 … how do you use artifact relics wow