Primary human dermal lymphatic endothelial cells (HDLECs, Promocell) were cultured according to the manufacturer’s protocol (Lonza) in EBM-2 medium containing EGM-2 MV SingleQuots. Cells of passage 5–6 were seeded in culture dishes at 80% confluence and transfected with miRNA control, miR-19b mimic, miRNA inhibitor control, or miR-19b inhibitor (GenePharma, China) using Lipofectamine 2000 (Invitrogen) for 24 h.
Mirna control
Manufactured by GenePharma
Sourced in China
The MiRNA controls are a set of synthetic RNA molecules designed to serve as positive and negative controls for microRNA (miRNA) analysis. These controls are used to validate the performance and accuracy of miRNA detection and quantification methods, such as real-time PCR, Northern blotting, or microarray analysis. The MiRNA controls provide a reliable and consistent standard for assessing the sensitivity, specificity, and reproducibility of miRNA-based experiments.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (5 mentions)
Mir 128 3p inhibitor, by GenePharma (2 mentions)
Psiren retroq vector, by Takara Bio (2 mentions)
Lipofectamine ltx, by Thermo Fisher Scientific (2 mentions)
Plus reagent, by Thermo Fisher Scientific (2 mentions)
Small interfering rnas sirna, by GenePharma (2 mentions)
Egm 2 mv singlequots, by Lonza (1 mentions)
Mirna inhibitor control, by GenePharma (1 mentions)
Hdlec, by Lonza (1 mentions)
Control vector, by GenePharma (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Mir 641 inhibitor, by GenePharma (1 mentions)
Sirnas, by GenePharma (1 mentions)
Small interfering rna, by GenePharma (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Wpmy 1, by American Type Culture Collection (1 mentions)
Control sirna sinc, by GenePharma (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Si lats2, by GenePharma (1 mentions)
11 protocols using mirna control
1
Transfection of HDLECs with miR-19b
2
Transfection of miRNA and siRNA
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The mimic and the inhibitor of hsa‐miR‐6817‐5p, miRNA control, circRFWD2 siRNA, circINO80 siRNA and control vector were synthesized by GenePharma Co. and shown in Table S1 . Cells were transfected by Lipofectamine 3000 Reagent (Invitrogen), when the cell density reached 80% confluency.
3
SNHG1 knockdown in cells
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SiRNAs targeting SNHG1, the miRNA control, miR-641, miR-641 inhibitory control, and miR-641 inhibitor were purchased by Gene Pharma (Shanghai, China). Si-NC (5′-UUCUCCGAACGUGUCACGUTT-3′), si-SNHG1-1 (5′-GAGCAAAUAAGGUGUAUAATT-3′), si-SNHG1-2 (5′-CCAGCACCUUCUAAATT-3′), and si-SNHG1-3 (5′-GGCCAUAGCUUUAAGAGAUTT-3′) served as control and si-SNHG1 sequences. The pcDNA-3.1-RRS1 plasmid was obtained from Shanghai GeneChem (China). Following the manufacturer's instructions, lipofectamine 2000 (Invitrogen) was used for cell transfection. sh-SNHG1 and negative control were synthesized by Obio (Shanghai, China).
4
Modulation of miR-128-3p and PVT1 by UPF1
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The pSIF–GFP–miR-128-3p precursor plasmid and the precursor control were gifts from Dr. Yong Li in Baylor College of Medicine. miR-128-3p inhibitors and miRNA controls were obtained from GenePharma Technology (Shanghai, China). Small interfering RNAs (siRNAs) targeting PVT1 and a negative control were also obtained from GenePharma Technology (Shanghai, China). Three shRNAs targeting human UPF1 and the shRNA negative control sequence were as follows: shUPF1#1, sense: 5’-GATCCGAGCCACATTGTAAATCATTTCAAGAGAATGATTTA CAATGTGGCTTTTTTTG-3, antisense: 5’-AATTCAAAAAAAGCCACATTGTAAATC ATTCTCTTGAAATGATTTACAATGTGGCTCG-3’; shUPF1#2, sense: 5’-GATCCG GCGAGAAGGACTTCATCATTCAAGAGATGATGAAGTCCTTCTCGCTTTTTTG-3’, antisense: 5’-AATTCAAAAAAGCGAGAAGGACTTCATCATCTCTTGAATGATGAA GTCCTTCTCGCCG-3’; shUPF1#3, sense: 5’-GATCCGGCAGCCACATTGTAAAT.
CATCAAGAGTGATTTACAATGTGGCTGCTTTTTTG-3’, antisense: 5’-AATTCA AAAAAGCAGCCACATTGTAAATCACTCTTGATGATTTACAATGTGGCTGCCG-3’. The shRNA constructs were cloned into Bam HI and Eco RI sites in an RNAi-ready pSIREN-RetroQ vector (Clontech, USA). All DNA constructs were confirmed by Sanger sequencing. Transfections were performed using Lipofectamine® LTX and Plus reagent (Invitrogen, USA) according to the manufacturer’s instructions.
CATCAAGAGTGATTTACAATGTGGCTGCTTTTTTG-3’, antisense: 5’-AATTCA AAAAAGCAGCCACATTGTAAATCACTCTTGATGATTTACAATGTGGCTGCCG-3’. The shRNA constructs were cloned into Bam HI and Eco RI sites in an RNAi-ready pSIREN-RetroQ vector (Clontech, USA). All DNA constructs were confirmed by Sanger sequencing. Transfections were performed using Lipofectamine® LTX and Plus reagent (Invitrogen, USA) according to the manufacturer’s instructions.
5
HOXA11-AS Overexpression in SCC-25 Cells
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Lipofectamine 2000 (Invitrogen, USA) was used to perform transfections according to the manufacturer’s instructions. miR-518a-3p mimics, miRNA controls, small interfering RNAs (siRNAs) specific to HOXA11-AS and a negative control siRNA were synthesized by GenePharma Technology (Shanghai, China). The HOXA11-AS overexpression plasmid, i.e. pcDNA3.1-HOXA11-AS, was acquired from Genearray Biotechnology (Shanghai, China), which was transfected to SCC-25 cells, followed by G418 selection. Briefly, the SCC-25 cells, which were seeded in 24-well plates at the density of 1 × 105 cells/cm2, were grown in complete medium containing 250 mg/mL G418 (Sigma-Aldrich, USA). After 48 h of transfection, cells were cultured for 4 weeks at 1:10 (v/v) in medium with 600 μg/mL G418. The resulting clones, termed as HOXA11-AS cells, were maintained in medium with 300 μg/mL G418. cloning qRT-qPCR was used to confirm HOXA11-AS overexpression in cells.
6
miR-497-5p Mimics and Inhibitors
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The human miR-497-5p mimics/inhibitor and their corresponding miRNA controls were synthesized by GenePharma (Shanghai, China). YAP1 and TEAD1 expression pcDNA3.1 plasmids were purchased from Origene (Beijing, China). Lipofectamine 2000 Reagent (Invitrogen, Carlsbad, CA, USA) was used for cell transfection according to the manufacturer’s instructions.
7
miR-128-3p Precursor Plasmid Protocol
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The pSIF-GFP-miR-128-3p precursor plasmid and the precursor control were gifts from Dr. Yong Li in Baylor College of Medicine. miR-128-3p inhibitors and miRNA controls were obtained from GenePharma Technology (Shanghai, China). Small interfering RNAs (siRNAs) targeting PVT1 and a negative control were also obtained from GenePharma Technology (Shanghai, China). Three shRNAs targeting human UPF1 and the shRNA negative control sequence were as follows: shUPF1#1, sense: 5'-GAT CCG AGC CAC ATT GTA AAT CAT TTC AAG AGA ATG ATTTA CAA TGT GGC TTT TTTTG-3, antisense:
5'-AAT TCA AAA AAA GCC ACA TTG TAA ATC ATT CTC TTG AAA TGA TTT ACA ATG TGG CTC G-3'; shUPF1#2, sense: 5'-GAT CCG GCG AGA AGG ACT TCA TCA TTC AAG AGA TGA TGA AGT CCT TCT CGC TTT TTTG-3' , antisense: 5'-AAT TCA AAA AAG CGA GAA GGA CTT CAT CAT CTC TTG AAT GAT GAA GTC CTT CTC GCC G-3'; shUPF1#3, sense: 5'-GAT CCG GCA GCC ACA TTG TAAAT.
CAT CAA GAG TGA TTT ACA ATG TGG CTG CTT TTTTG-3' , antisense: 5'-AAT TCA AAA AAG CAG CCA CAT TGT AAA TCA CTC TTG ATG ATT TAC AAT GTG GCT GCCG-3' . The shRNA constructs were cloned into Bam HI and Eco RI sites in an RNAi-ready pSIREN-RetroQ vector (Clontech, USA). All DNA constructs were confirmed by Sanger sequencing. Transfections were performed using Lipofectamine ® LTX and Plus reagent (Invitrogen, USA) according to the manufacturer's instructions.
5'-AAT TCA AAA AAA GCC ACA TTG TAA ATC ATT CTC TTG AAA TGA TTT ACA ATG TGG CTC G-3'; shUPF1#2, sense: 5'-GAT CCG GCG AGA AGG ACT TCA TCA TTC AAG AGA TGA TGA AGT CCT TCT CGC TTT TTTG-3' , antisense: 5'-AAT TCA AAA AAG CGA GAA GGA CTT CAT CAT CTC TTG AAT GAT GAA GTC CTT CTC GCC G-3'; shUPF1#3, sense: 5'-GAT CCG GCA GCC ACA TTG TAAAT.
CAT CAA GAG TGA TTT ACA ATG TGG CTG CTT TTTTG-3' , antisense: 5'-AAT TCA AAA AAG CAG CCA CAT TGT AAA TCA CTC TTG ATG ATT TAC AAT GTG GCT GCCG-3' . The shRNA constructs were cloned into Bam HI and Eco RI sites in an RNAi-ready pSIREN-RetroQ vector (Clontech, USA). All DNA constructs were confirmed by Sanger sequencing. Transfections were performed using Lipofectamine ® LTX and Plus reagent (Invitrogen, USA) according to the manufacturer's instructions.
8
Transfection and Silencing of MCM3AP-AS1 in Prostate Cancer
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Human PCa cell lines (PC-3, DU145, 22RV1, LNCaP) and normal follicular epithelial cell line (WPMY-1) were provided by ATCC, USA. Cells were incubated in RPMI 1640 with 10% FBS at 37°C. Control siRNA (si-NC), si-MCM3AP-AS1, miR-543-3p mimics, miR-543-3p inhibitor, pcDNA3.1, pcDNA3.1- MCM3AP-AS1, control miRNA, lentivirus-sh-MCM3AP-AS1 (LV-sh-MCM3AP-AS1) and control lentivirus-sh (LV-shRNA-NC) were made by GenePharma, China, and transfected to cells using Lipofectamine 2000 (Invitrogen, USA).
9
LINC00174 regulation in lung cancer
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pcDNA3.1 vectors containing the LINC00174 sequence (pcDNA3.1-LINC00174), empty vector,
small interfering RNA (siRNA) targeting LATS2 (si-LATS2), scramble siRNA negative control
(si-NC), miR-31-5p mimic (5ˊ-CUUUUUGCGGUCUGGGCUUGC-3ˊ), miR-31- 5p inhibitor
(5ˊ-CGUUCGGGUCUGGCGUUUUC-3ˊ) and the control miRNA (5ˊ-UCACAACCUCCUAGAAAGAGUAGA-3ˊ) were
purchased from Shanghai GenePharma Co., Ltd. A549 and H1299 cells were respectively
transfected with Lipofectamine® 2000 (Invitrogen, Thermo Fisher Scientific,
Inc., USA) as instructions.
small interfering RNA (siRNA) targeting LATS2 (si-LATS2), scramble siRNA negative control
(si-NC), miR-31-5p mimic (5ˊ-CUUUUUGCGGUCUGGGCUUGC-3ˊ), miR-31- 5p inhibitor
(5ˊ-CGUUCGGGUCUGGCGUUUUC-3ˊ) and the control miRNA (5ˊ-UCACAACCUCCUAGAAAGAGUAGA-3ˊ) were
purchased from Shanghai GenePharma Co., Ltd. A549 and H1299 cells were respectively
transfected with Lipofectamine® 2000 (Invitrogen, Thermo Fisher Scientific,
Inc., USA) as instructions.
10
Establishing Stable miRNA Expression
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Cervical cancer cells (2 × 105 cells) were transfected with 1 μg of a plasmid expressing miR-486-5p inhibitor sponge, miR-486-5p mimic, or control miRNA (Genepharma, Shanghai, China). Transfection into cells was carried out according to the manufacturer’s instructions using Lipofectamine3000 for 48 h (Invitrogen–Life Technologies). Stable expression of each miRNA was established in the cells after 15 days of incubation in complete DMEM with blasticidin (12 μg/mL). We verified miRNA expression in the clones using real-time PCR and integrate all the successful clones for the subsequent experiments.
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