The biotin‐coupled circ_0008194 probe and control oligo probe were designed and synthesized by RiboBio (Guangzhou). Briefly, cells were lysed with lysis buffer and 50 μl of the cell lysates were aliquot for input. After incubated with the biotin‐coupled probe at room temperature for 1 h, the rest were incubated with streptavidin magnetic beads (Life Technologies) at 4°C for 4 h to capture the probe‐RNA complexes. Then, the beads were washed with lysis buffer. Finally, RNAs bound to the beads were isolated with TRIzol Reagent (Invitrogen) and analyzed by qRT‐PCR.
Control oligo probe
Manufactured by RiboBio
Sourced in China
The Control oligo probe is a laboratory tool used to assess the performance and reliability of nucleic acid detection assays. It serves as a reference standard to ensure the proper functioning of the experimental setup. The probe provides a consistent and known target sequence that can be used to validate the specificity and sensitivity of the detection system.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using control oligo probe
1
Biotin-coupled Circular RNA Pulldown Assay
2
Biotin-coupled miR-498 Probe Enrichment
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The biotin-coupled miR-498 probe and control oligo probe were synthesized by RiboBio (Guangzhou, China). Briefly, HCC827/OTR and H1975/OTR cells were lysed using lysis buffer. Cell lysates were incubated with the biotin-coupled miR-498 probe or biotin-coupled NC probe for 1 h at room temperature, and then the remaining cell lysates were cultured with streptavidin magnetic beads (Life Technologies, Mountain View, CA, USA). Subsequently, the lysis buffer was used to wash the beads. Finally, RNAs bound to the beads were extracted using TRIzol Reagent (Invitrogen), and then analyzed using qRT-PCR.
3
Biotin-Coupled circFAM73A Probe Capture
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The biotin-coupled cricFAM73A probe, control oligo probe, miR-490-3p wild-type probe, and mutated-type probe were synthesized by RiboBio (Guangzhou, China). Cells were fixed by 1% formaldehyde for 10 min and lysed with lysis buffer containing 25 mM Tris-HCl pH 7.4, 150 mM NaCl, 1 mM EDTA, 1% NP-40, 5% glycerol, 1 U/μL SUPERase-In RNase Inhibitor (Ambion), and Protease Inhibitor Cocktail (Halt). The lysates were then centrifuged at 10,000 g for 10 min. Fifty microliter of the supernatant was saved for input analysis and the remaining part was transferred to a fresh tube. The biotin-coupled RNA complex was pulled down by incubating streptavidin-coated magnetic beads (Invitrogen, Carlsbad, USA) with cell lysates overnight at 4 °C. On the next day, the beads-probes-RNA mixture was washed and the RNA bound to the beads was extracted by TRIzol for subsequent qRT-PCR analysis.
The relative abundance of RNA bound to the probe was analyzed using 2-ΔCt method relative to the RNA levels of input. The miRNA expression bound to circFMA73A was then calculated by the expression of miRNA captured by the circFAM73A probe relative to miRNA captured by the Oligo probe.
The relative abundance of RNA bound to the probe was analyzed using 2-ΔCt method relative to the RNA levels of input. The miRNA expression bound to circFMA73A was then calculated by the expression of miRNA captured by the circFAM73A probe relative to miRNA captured by the Oligo probe.
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