Mir-125b mimic, miR-125b inhibitor, and the miRNA NC (negative control) mimic and inhibitor were synthesized by RiBoBio (Guangzhou, China). The STAT3 and negative control siRNA were purchased from Sigma-Aldrich. MiRNA or siRNA were transfected into cells according to the manufacturer’s instructions using Oligofectamine™ as the transfection reagent (Life Technologies, CA, USA). Cells were harvested at 72 h after transfection and then used for experiments.
Mir 125b mimic
Manufactured by RiboBio
Sourced in China
The Mir-125b mimic is a synthetic RNA molecule designed to mimic the function of the naturally occurring microRNA miR-125b. MicroRNAs are small non-coding RNAs that play a crucial role in regulating gene expression. The Mir-125b mimic can be used for research purposes to study the biological functions and potential applications of miR-125b in various cellular and molecular processes.
Automatically generated - may contain errors
Lab products found in correlation
Mir 125b inhibitor, by RiboBio (3 mentions)
Oligofectamine, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Brd4 small interfering rna sirna, by RiboBio (1 mentions)
Oligofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Sybr premix ex taq 2, by Takara Bio (1 mentions)
Control sirna, by RiboBio (1 mentions)
Primescript rt reagent kit with gdna eraser, by Takara Bio (1 mentions)
Control mirna, by RiboBio (1 mentions)
Stat3 sirna, by RiboBio (1 mentions)
Rnaiso plus, by Takara Bio (1 mentions)
E coli poly a polymerase, by New England Biolabs (1 mentions)
7500 real time pcr system, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Nucleofector transfection kit, by Lonza (1 mentions)
Cancer cell isolation kit, by Panomics (1 mentions)
5 protocols using mir 125b mimic
1
miRNA Transfection Protocol for STAT3
2
miR-125b Modulation in Cell Transfection
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The cells (50% confluence) were transfected with miR-125b mimic (50, 100 and 150 nmol/l), inhibitor (25, 50 and 100 nmol/l) or BRD4 small interfering RNA (siRNA; 50 nmol/l; all Guangzhou RiboBio Co., Ltd., Guangzhou, China), according to the manufacturer's protocols. For experiments where only one concentration was used, cells were transfected with 50 nmol/l mimic/inhibitor/siRNA. miR-negative control (NC; 50 nmol/l), inhibitor NC (50 nmol/l) and si-NC (50 nmol/l) were used as respective transfection controls. The transfection reagent used was Lipofectamine® 2000 (Thermo Fisher Scientific, Inc.). The sequence of BRD4 siRNA was 5′-GACUAGAAACUUCCCAAAUGUCUUUCAAGAGAAGACAUUUGGGAAGUUUCUAGUC-3′. The sequence of miR-125b mimic was sense, 5′-UCACAAGUUAGGGUCUCAGGGA-3′ and antisense, 3′-AGUGUUCAAUCCCAGAGUCCCU-5′. The sequence of miR-125b inhibitor was 5′-UCACAAGUUAGGGUCUCAGGGA-3′. The sequence of miR-NC was 5′-GGCUGCCUACUUAGCUUGAGAGUG-3′. The sequence of inhibitor NC was 5′-ACGGGUGUGACCACUCCAGGCUGC-3′. The sequence of si-NC was 5′-CCAUCUCCCGGUACAAAAUCUGCU-3′. At 48 h following transfection, the cells were used for protein extraction.
3
Upregulation of miR-125b in GBC cells
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Human miR-125b mimics oligonucleotides (miR-125bmimic) and control non-specific miRNA mimics oligonucleotides (C-miR) were purchased from RiboBio (Guangzhou, China). GBC cell lines, TYGBK-8 and G-415 cells, were transfected with C-miR or miR-125b-mimic with Oligofectamine 2000 (Thermo Fisher Scientific, USA) for 48 h. Forty-eight hours after transfection, endogenous miR-125b expression was examined by qRT-PCR to confirm the upregulation efficiency.
4
miR-125b Expression Analysis in Skin Conditions
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Total RNA was isolated from HaCaT cells, cholesteatoma, and the corresponding skin with RNAiso plus (TaKaRa Biotechnology Co. Ltd., Dalian, China). The integrity, purity, and concentration of total RNA were analysed by spectrophotometry and gel electrophoresis. Then, the RNA was polyadenylated using E. coli Poly (A) Polymerase (New England Biolabs, MA) and reverse-transcribed into cDNA with a PrimeScriptTM RT reagent Kit with gDNA Eraser (TaKaRa). Finally, quantitative real-time PCR was carried out using a SYBR®Premix Ex Taq™II (TaKaRa) and 7500 real-time PCR system (Applied Biosystems, USA). The relative expression of gene levels was calculated using the 2–ΔΔCT method. U6 served as an internal control normalised the expression data of miR-125b. The sequences of the primers: miR-125b-5p: 5’-TCCCTGAGACCCTAACTTGTGA-3’ miR-reverse: n5’-GCTGTCAACGATACGCTACG-3’; miR-RT primer: 5’-GCTGTCAACGATACGCTACGTAACGGCATGACAGTGTTTTTTTTTTTTTTTTTTTTTTT-3’; U6 forward: 5’-CTCGCTTCGGCAGCACA-3’ and reverse: 5’-AACGCTTCACGAATTTGCGT-3’; HaCaT cells were transiently transfected with miR-125b mimics, miR-125b inhibitor, control miRNA, STAT3 siRNA, and control siRNA (all from RiboBio Co. Ltd., Guangzhou, China), using Lipofectamine® 3000 reagent (Invitrogen, Carlsbad, CA, USA).
5
Isolation and Culture of NSCLC Cells
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NSCLC cells were isolated from surgical tumor tissues using Cancer Cell Isolation Kit (Panomics). According to the manufacture’s instructions, cells were cultured for 4 days at 37 °C under 5 % CO2 in complete RPMI 1640 medium (GIBCO, containing 10 % heat-inactivated fetal bovine serum supplemented with 2 mM glutamine, 100 IU/ml penicillin and 100 mg/ml streptomycin sulfate), and used for experimental research. MiR-125b mimics and miR-125b inhibitor were from Ribobio (Guangzhou, China). Human TP53INP1 expression plasmid was purchased from Origen. Human TP53INP1 siRNA was from Santa Cruz. The nucleofector transfection kit was purchased from Amaxa.
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