The biotinylated (bio)-NC and bio-miR-495-3p were acquired from Sigma Aldrich. In short, 22Rv1 and LNCaP cells were transfected with bio-NC or bio-miR-495-3p, respectively. Then, the cells were lysed with lysis buffer. Next, the lysate was incubated with streptavidin-coupled magnetic beads for 2 h. Thereafter, the bound RNA was obtained via using Trizol reagent, and the level of TRIP13 was determined via qRT-PCR.
Bio nc
Manufactured by Merck Group
Sourced in United States
The Bio)-NC is a lab equipment product designed for scientific research and analysis. It serves as a core function in various laboratory applications, but without further details on its intended use, a more specific description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
4 protocols using bio nc
1
Biotinylated miR-495-3p Pulldown Assay
2
MiR-130a-3p Target Identification Protocol
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Biotin-labeled MiR-130a-3p (Bio-miR-130a-3p) and miR-negative control (Bio-NC) were designed and synthesized by Thermo Fisher (USA). MKN45 and AGS cells were lysed and subsequently incubated with Bio-miR-130a-3p or Bio-NC for 2 h. Next, the suspension was incubated overnight with streptavidin beads (Sigma, USA) at 4°C. Beads were washed and the elution was purified with an RNA purification Kit (Cat#: DP412, Tiangen, China). After the enrichment, GCNT4 was quantified by RT-qPCR [26 (link)].
3
RNA Pulldown Assay for circNOL10
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For the RNA pulldown assay, SW620 and SW480 cells were infected with 3ʹ-biotinylated miR-135a-5p mimic or miR-135b-5p (Millipore). After transfection 48 h, approximately 1 × 107 cells were lysed and incubated with streptavidin-coupled beads at room temperature for 2 h to form biotin-miRNA-lncRNA complexes, while bio-NC (Millipore) was used as a negative control. After that, RNA was extracted and purified with Trizol reagent and proteinase K, respectively. The abundance of circNOL10 was measured with RT-qPCR assay.
4
Quantification of miR-124-3p via Biotin-NEAT1
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According to the manufacturer’s instruction, biotin-labeled NEAT1 (Bio-NEAT1) or Bio-NC (Millipore, Bedford, MA, USA) was respectively transfected into cells. 48 h later, the cell lysate was incubated with streptavidin agarose-magnetic beads at 4°C for 1 h. Ultimately, the miR-124-3p content in bead-RNA complex was probed by qRT-PCR.
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