Biotin-labeled circMID1 probe, the wild-type (wt)/mutate-type (mut) of miR-330-3p probe (bio-miR-330-3p-wt/mut) and their negative control probes (NC probe and bio-NC) were synthesized by Genepharma. PC-3 and DU145 cells were harvested, and then the cell lysates were incubated with Dynabeads M-280 Streptavidin (Invitrogen) coated circMID1 probe or NC probe. The enrichment of candidate miRNAs was examined by qRT-PCR to screen the targeted miRNA for circMID1. To explore whether miR-330-3p could pull down circMID1, YTHDC2 and IGF1R, PCa cells were transfected with the bio-miR-330-3p-wt/mut probe or bio-NC probe for 24 h, as the previous study [16 (link)]. Then, the cell lysates were cultured with the Dynabeads, and the RNA enrichment was analyzed by qRT-PCR.
Bio nc
Manufactured by GenePharma
Sourced in China
The Bio-NC is a laboratory equipment designed for nucleic acid separation and purification. It utilizes a nanocellulose-based membrane to effectively capture and isolate DNA, RNA, or other nucleic acid molecules from complex samples.
Automatically generated - may contain errors
Lab products found in correlation
Dynabeads m 280 streptavidin, by Thermo Fisher Scientific (14 mentions)
Biotin rna labeling mix, by Roche (5 mentions)
M 280 streptavidin magnetic bead, by Thermo Fisher Scientific (5 mentions)
Rna structure buffer, by Thermo Fisher Scientific (4 mentions)
Bio nc, by Thermo Fisher Scientific (4 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
Dynabeads m 280 streptavidin beads, by Thermo Fisher Scientific (2 mentions)
Bio malat1 wt, by GenePharma (2 mentions)
Bio malat1 mut, by GenePharma (2 mentions)
Streptavidin agarose bead, by Thermo Fisher Scientific (2 mentions)
Streptavidin coated magnetic beads, by Thermo Fisher Scientific (2 mentions)
Biotin labeled circ 0002060 probe bio circ 0002060, by GenePharma (2 mentions)
M 280 streptavidin, by Merck Group (1 mentions)
Rnasin, by Promega (1 mentions)
Streptavidin dynabeads, by Thermo Fisher Scientific (1 mentions)
M 280 streptavidin magnetic beads, by Merck Group (1 mentions)
Magnetic bead, by Merck Group (1 mentions)
Ripa lysis buffer, by Thermo Fisher Scientific (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Sirna mate transfection reagent, by GenePharma (1 mentions)
38 protocols using bio nc
1
Identifying circMID1-Targeting miRNAs in Prostate Cancer
2
Biotin-labeled circFBXW7 Pulldown Assay
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Probe-control or probe-circFBXW7 from sh-circFBXW7 vector was transcribed and labeled by Biotin RNA Labeling Mix (Roche, Basel, Switzerland). An RNA structure buffer (Thermo, MA, USA) was used to induce secondary structure formation from the biotin-labeled RNAs. The biotinylated circFBXW7 and negative control (bio-NC) were generated via GenePharma and coated to streptavidin-conjugated magnetic beads. Magnetic beads were applied to incubate the cells for 6 h after cells were lysed. The RNA on the beads was isolated and RT-qPCR was applied to assess the enrichment of miR-494-3p.
3
Investigating NR2F2-AS1 Binding to miR-4429
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Pull-down assay was utilized to examine the underlying binding capacity between NR2F2-AS1 with miR-4429. NR2F2-AS1-Wt, NR2F2-AS1-Mut, and NC were biotinylated to be Bio-NR2F2-AS1-Wt, Bio-NR2F2-AS1-Mut, and Bio-NC by GenePharma Company (Shanghai, China). Bio-NR2F2-AS1-Mt, Bio-NR2F2-AS1-Mut and Bio-NC were transfected into HeLa or SiHa cells. After incubation of 48 h, cells were lysed, and the cell lysate was cultured with Dynabeads M-280 Streptavidin (Invitrogen, CA). Purified RNA complex was detected by qRT-PCR.
4
Investigating LINC00346 in Colorectal Cancer
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Bio-LINC00346-Wt, Bio-LINC00346-Mut and Bio-NC (GenePharma) were transfected into CRC cells (HT29 and HCT116) using Lipofectamine 3000 (Invitrogen). After cultured for 48 h, cells were incubated with Dynabeads M-280 Streptavidin beads (Invitrogen) for 1 h. The RNAs were detected by qRT-PCR.
5
Biotin-labeled MALAT1 Enrichment
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Bio-MALAT1-Wt, Bio-MALAT1-Mut, and Bio-NC were obtained from GenePharma (Shanghai, China). After 48 h of transfection with Bio-MALAT1-Wt, Bio-MALAT1-Mut, and Bio-NC, the cells were incubated with Dynabeads M-280 Streptavidin (Invitrogen, USA). RT-qPCR was used to examined the enrichment of miR-206.
6
Biotinylated miRNA-mRNA Binding Assay
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Bio-LMCD1-AS1-Wt, Bio-LMCD1-AS1-Mut and Bio-NC were synthesized by GenePharma. The biotinylated miRNA or mRNA was co-incubated with cell lysates overnight. Next, the magnetic beads with streptavidin (Invitrogen) were added and cultured for 48 h. Finally, the purified RNA complex was detected by qRT-PCR.
7
Biotin-labeled miRNA Interactome Identification
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Three diverse biotin-labeled miRNA sequences WT miR-24-3p (Bio-miR-24-3p-WT), MUT miR-24-3p (Bio-miR-24-3p-MUT, sequence mutation that complementary to MCM3AP-AS1), and a random miRNA that did not complementary to MCM3AP-AS1 (Bio-NC) were designed and synthesized by GenePharma. The miRNAs were transfected into VECs for 48 h when the cell confluence reached 80–90%. Subsequently, cells were lysed using lysis buffer and the lysate was co-cultured with magnetic beads coated with M-280 streptavidin (Sigma-Aldrich Chemical Company, MO, USA) at 4°C for 3 h. The beads were rinsed and protein-nucleotide complex absorbed by the beads were eluted. Trizol was used to extract the total RNA, and MCM3AP-AS1 expression was assessed using RT-qPCR.
8
Bio-DLEU2 Pulldown Assay
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DLEU2 and negative control (NC) were biotinylated to be bio-DLEU2, and bio-NC by GenePharma Company (Shanghai, China). Next, bio-DLEU2 or bio-NC was incubated with Dynabeads M-280 Streptavidin (Invitrogen, USA) for 1 hour at 4 °C. Then, Hela and SiHa cells were dissolved in the soft lysis buffer plus 80 U/mL RNasin (Promega Madison, WI, USA) and the cell lysates were incubated with the RNA-bound beads. Finally, the beads were washed with buffer and miR-128-3p were quantified and analyzed by qPCR.
9
Biotinylated miR-378a-3p Pulldown Assay
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The biotinylated miR-378a-3p Wt/Mut (bio-miR-378a-3p Wt/Mut) and the negative control (bio-NC) were purchased from GenePharma (Shanghai, China) and were transfected into SGC7901 and AGS cells. After 24 h of transfection, the cells were lysed and collected. After incubation with Streptavidin agarose beads (Invitrogen) for 10 min, RT-qPCR was applied to determine the enrichment of ACTA2-AS1. The assay was performed according to previous studies [40 (link)].
10
Biotinylated miR-137 Pulldown Assay
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For pull-down assay, miR-137 without complementary sites with Notch1 was seen as internal reference (termed NC). MiR-137, miR-137-Mut, and NC were labeled with biotin to generate Bio-miR-137, BiomiR-137-Mut, and Bio-NC by GenePharma Company (Shanghai, China). Thereafter, they were transfected into CD14 + PBMCs. Forty-eight hours later, cells were harvested and incubated with Dynabeads M-280 Streptavidin (Invitrogen) for 10 min. After washing three times with buffer solution, the enrichments of bound RNAs were quantified and analyzed by qRT-PCR.
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