The MiRNA-610 agonist (agomir-610) and negative control (agomir-NC) were purchased from Thermo Fisher Scientific (Foster City, CA, USA). The expression vector (pc-HDGF) and negative control vector (pc-NC) were constructed by Genscript (Nanjing, China). 50 nM agomir-610 or agomir-NC and 1.5 μg pc-HDGF or pc-NC were transfected with LipofectamineTM 3000 Reagent (Invitrogen, Foster City, CA, USA) according to the operating manual. After 4 h, normal media was applied to cultured cells with 48 h.
Agomir nc
Manufactured by Thermo Fisher Scientific
Sourced in United States
AgomiR-NC is a type of laboratory equipment used in scientific research. It serves as a negative control for experiments involving microRNA (miRNA) analysis. The core function of AgomiR-NC is to provide a baseline reference for comparison in miRNA-related studies, without any specific intended effects or applications.
Automatically generated - may contain errors
Lab products found in correlation
Antagomir nc, by Thermo Fisher Scientific (6 mentions)
Agomir nc, by RiboBio (4 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Antagomir nc, by GenePharma (2 mentions)
Agomir nc, by GenePharma (2 mentions)
Si nc, by Thermo Fisher Scientific (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Antagomir nc, by RiboBio (2 mentions)
Pc nc, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions)
Polybrene, by Santa Cruz Biotechnology (1 mentions)
Dars as1 sirna1, by RiboBio (1 mentions)
Nc sirna, by Thermo Fisher Scientific (1 mentions)
Puromycin, by Thermo Fisher Scientific (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Ki8751, by MedChemExpress (1 mentions)
Pmir rb report vector, by RiboBio (1 mentions)
Dual luciferase reporter assay system, by Promega (1 mentions)
Si nc, by RiboBio (1 mentions)
Antagomir, by RiboBio (1 mentions)
11 protocols using agomir nc
1
MiRNA-610 Agonist Transfection Protocol
2
Transfection and Overexpression of DARS-AS1 and HMGB1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DARS-AS1 siRNA1, DARS-AS1 siRNA2, DARS-AS1 siRNA3, siRNA negative control (siRNA-NC), miR-188-5p agomir and miR-188-5p agomir negative control (agomir-NC) were purchased from Guangzhou RiboBio Co., Ltd (Guangzhou, China). The DARS-AS1 siRNA1, DARS-AS1 siRNA2, DARS-AS1 siRNA3, siRNA-NC, miR-188-5p agomir and agomir-NC were transfected into SiHa or HeLa cells using Lipofectamine 2000 (Thermo Fisher Scientific) for 24 h according to the manufacturer’s protocols.
The HMGB1-overexpressing and control lentiviruses were obtained from Ribobio (Guangdong, China), named HMGB1-OE and control vector (Ctrl-vector). 293 T cells were transfected with lentivirus, packaging plasmid (pAX2) and envelope plasmid (pMD2.G) for 72 h. After that, virus-containing supernatant was collected, and added into SiHa and HeLa cells in the presence of polybrene (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 48 h. Later on, cells were cultured with 2.5 μg/mL puromycin (Thermo Fisher Scientific) to select stable HMGB1-OE cells. Western blot assay was performed to verify the transfection efficiency of the lentiviruses.
The HMGB1-overexpressing and control lentiviruses were obtained from Ribobio (Guangdong, China), named HMGB1-OE and control vector (Ctrl-vector). 293 T cells were transfected with lentivirus, packaging plasmid (pAX2) and envelope plasmid (pMD2.G) for 72 h. After that, virus-containing supernatant was collected, and added into SiHa and HeLa cells in the presence of polybrene (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 48 h. Later on, cells were cultured with 2.5 μg/mL puromycin (Thermo Fisher Scientific) to select stable HMGB1-OE cells. Western blot assay was performed to verify the transfection efficiency of the lentiviruses.
3
Modulating Atherosclerosis with miRNA Agonists
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
micrON miRNA agomir-miR-19a, agomir-miR-19b, and agomir-NC were purchased from RiboBio (RiboBio, Guangdong, China). Agomir-miRNA and agomir-NC transfection was performed using the Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions.
Sequences were as follows. hsa-miR-19a: agomir sense: 5′-AGUUUUGCAUAGUUGCACUACA-3′; antisense: 5′-AsCs GUGCAACUAUGCAAAA AsAsUsUs-Chol-3′. hsa-miR-19b: agomir sense: 5′-AGUUUUGCAGGUUUGCAUCCAGC-3′; antisense: 5′-AsCs GAUGCAAACCUGCAAAA AsAsUsUs-Chol-3′.
Normal saline was mixed with agomir-miR-19a, agomir-miR-19b, and agomir-NC. The mice that successfully established atherosclerosis models were randomly divided into an ApoE−/− mice group, ApoE−/−+agomir-miR-19a/b-treated mice, and an agomir-NC mice group, with 10 mice in each group. The ApoE−/−+agomir-miR-19a/b-treated mice group was injected with a mixture of 5 nmol agomir-miR-19a/b through the tail vein, the NC mice group was injected with a mixture of 5 nmol agomir-NC through the tail vein, and the ApoE−/− mice group was injected with the same amount of agomir-NC through the tail vein every 3 days for 3 weeks. RT-qPCR was used to detect whether miR-19a and miR-19b were overexpressed in ApoE−/−+agomir-miR-19a/b-treated mice.
Sequences were as follows. hsa-miR-19a: agomir sense: 5′-AGUUUUGCAUAGUUGCACUACA-3′; antisense: 5′-AsCs GUGCAACUAUGCAAAA AsAsUsUs-Chol-3′. hsa-miR-19b: agomir sense: 5′-AGUUUUGCAGGUUUGCAUCCAGC-3′; antisense: 5′-AsCs GAUGCAAACCUGCAAAA AsAsUsUs-Chol-3′.
Normal saline was mixed with agomir-miR-19a, agomir-miR-19b, and agomir-NC. The mice that successfully established atherosclerosis models were randomly divided into an ApoE−/− mice group, ApoE−/−+agomir-miR-19a/b-treated mice, and an agomir-NC mice group, with 10 mice in each group. The ApoE−/−+agomir-miR-19a/b-treated mice group was injected with a mixture of 5 nmol agomir-miR-19a/b through the tail vein, the NC mice group was injected with a mixture of 5 nmol agomir-NC through the tail vein, and the ApoE−/− mice group was injected with the same amount of agomir-NC through the tail vein every 3 days for 3 weeks. RT-qPCR was used to detect whether miR-19a and miR-19b were overexpressed in ApoE−/−+agomir-miR-19a/b-treated mice.
4
Manipulating ZFAS1, SP1, and miR-150-5p in Colorectal Cancer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ZFAS1 siRNA lentivirus (siZFAS1-1, siZFAS1-2, siZFAS1-3, siZFAS-4) were obtained from Applied Biologic Materials, Inc (Canada) to knock down the expression of ZFAS1 and a scramble siRNA GFP Lentivector (scramble) was performed as a control, the siRNA sequence was listed in Supplementary materials and methods. Two CRC cell lines HCT116 and HCT8 were infected and stable cells were selected by treatment of puromycin (10 µg/ml). Small interferece RNA (siSP1-1, siSP1-2) targeting SP1 were chemically synthesized (Applied Biologic Materials, Inc, Canada), and the squence was listed in Supplementary materials and methods.
AgomiR-150-5p (miR-150-5p mimic), antagomiR-150–5p (anti-miR-150-5p) and their corresponding negative control (agomiR-NC, antagomiR-NC) were obtained from GenePharma (Shanghai, China), and their sequences were listed inSupplementary materials and methods. The agomiR-150-5p and agomiR-NC, antagomiR-150-5p and antagomiR-NC, siSP1, siSP2 and scramble, pcDNA3.1-VEGFA, pcDNA3.1-ZFAS1, pcDNA3.1-SP1, and blank vector (vector) were transfected into cells using LipofectamineTM 2000 (Invitrogen, USA) following the manufacturer’s protocol. Additionally, cell without transfection were treated with a VEGFR2 inhibitor Ki8751 (40 nM, Medchem Express, USA).
AgomiR-150-5p (miR-150-5p mimic), antagomiR-150–5p (anti-miR-150-5p) and their corresponding negative control (agomiR-NC, antagomiR-NC) were obtained from GenePharma (Shanghai, China), and their sequences were listed in
5
Validating miR-24 Regulation of S100A8
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bioinformatic software, including miRanda and TargetScan, was used to predict that miR-24 could specifically combine with the 3′ untranslated region (3′UTR) of S100A8 gene. The wild-type S100A8 3′UTR binding site and mutant binding site were cloned into pmiR-RB-REPORT™ vector (RiboBio, Guangzhou, China) to construct wild-type luciferase reporter vector (pmiR-S100A8-wt) and mutant-type vector (pmiR-S100A8-mut) by Genescript (Nanjing, China). In this assay, the luciferase reporter vector, miR-24 agonist (agomiR-24), or negative control agonist (agomiR-NC) (Invitrogen, Foster City, CA) were cotransfected into HEK 293T cells. After that, the dual-luciferase reporter assay system (Promega, Madison, WI) was used to check the relative luciferase activity according to the manufacturer’s protocol.
6
Targeting LINC00958 and miR-625 in Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The small interfering RNA (siRNA) that targeted LINC00958 (si-LINC00958) and negative control siRNA (si-NC) were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China). Agomir-625, agomir-NC, antagomir-625, and antagomir-NC were constructed at Shanghai GenePharma Co., Ltd. (Shanghai, China). The empty pcDNA3.1 plasmid and NUAK1-overexpressing plasmid pcDNA3.1-NUAK1 (pc-NUAK1) were chemically synthesized by the Chinese Academy of Sciences (Changchun, China).
Cells were seeded in 6-well plates with the culture medium without antibiotics one night before transfection. To obtain cell models with high or low expression of a target gene, the si-LINC00958 (100 pmol), si-NC (100 pmol), agomir-625 (50 nM), agomir-NC (50 nM), antagomir-625 (100 nM), antagomir-NC (100 nM) and plasmids (4 μg) were transfected into cells using the Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA). After different periods of incubation, the transfected cells were collected for the subsequent functional assays.
Cells were seeded in 6-well plates with the culture medium without antibiotics one night before transfection. To obtain cell models with high or low expression of a target gene, the si-LINC00958 (100 pmol), si-NC (100 pmol), agomir-625 (50 nM), agomir-NC (50 nM), antagomir-625 (100 nM), antagomir-NC (100 nM) and plasmids (4 μg) were transfected into cells using the Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA). After different periods of incubation, the transfected cells were collected for the subsequent functional assays.
7
Modulation of miR-204-5p in hADSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mimics, mimic-NC, agomir (cholesterol-conjugated 2′-O-methyl-modified mimics), agomir-NC, antagomir and antagomir-NC of miR-204-5p, were synthe-sized by Guangzhou RiboBio Co., Ltd. (Guangzhou, China). The miR-204-5p target site was predicted using online software, and found to be highly conserved among vertebrates. pVAX1- DVL3 and pVAX1-NC were purchased from Biovector Science Lab, Inc. (Beijing, China), and siRNA-DVL3 (si-DVL3) and siRNA-NC (si-NC) were prepared by Invitrogen (Carlsbad, CA, USA). The hADSCs were transfected with mimics, mimic-NC, agomir, agomir-NC (100 nM), antagomir, antagomir-NC (200 nM), pVAX1-DVL3, pVAX1-NC, siRNA-DVL3 or siRNA-NC using Lipofectamine 2000 (Invitrogen) according to the standard manufacturer-recommended protocol. At 2 days following transfection, cell differentiation was induced using the aforementioned protocol.
8
MiR-183-5p Regulation in VSMCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human VSMCs were purchased from the American Type Culture Collection (ATCC). Cells were grown in the Dulbecco's modi ed Eagle's medium (DMEM; Hyclone, GE Health Care, USA) with 10% fetal bovine serum (FBS; Gibco, Thermo Fisher Scienti c, Waltham, USA). Then cells were incubated in a humidi ed atmosphere with 5% CO 2 at 37℃. MiR-183-5p agomir, miR-183-5p antagomir, and their relative control miRNAs (agomir NC and antagomir NC) were provided by Ribobio (Guangzhou, China). The sequences were as follows: miR-183-5p agomirs, 5'-UAUGGCACUGGUAGAAUUCACU-3'; miR-183-5p antagomir, 5'-AGUGAAUUCUACCAGUGCCAUA-3'; agomirs NC, 5'-UGUAGGGCCACUCAGUCAACUU-3'; antagomir NC, 5'-AAUAUGGGCGAAAUGGGGCCAUC-3'.
As previous study described, cells were transfected with 100 nM miR-183-5p agomir or antagomir, or agomir NC, antagomir NC to regulate the expression level of miR-183-5p by using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions [18] . 48h post-transfection, cells were collected for further experiments. To investigate ox-LDL regulation on miR-183-5p expression, VSMCs were treated with ox-LDL (40 μg/ml).
As previous study described, cells were transfected with 100 nM miR-183-5p agomir or antagomir, or agomir NC, antagomir NC to regulate the expression level of miR-183-5p by using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions [18] . 48h post-transfection, cells were collected for further experiments. To investigate ox-LDL regulation on miR-183-5p expression, VSMCs were treated with ox-LDL (40 μg/ml).
9
Regulation of OIP5-AS1 and miR-143-3p
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The si-OIP5-AS1, si-NC, pcDNA3.1-OIP5-AS1, pcDNA3.1-NC, miR-143-3p antagomir, NC antagomir, miR-143-3p agomir and NC agomir were designed and synthesized by Invitrogen (Shanghai, China). Twenty-four h before transfection, cells were aspirated into a six-well plate (1 × 105 cells/well) with fresh serum-free medium. Following transfection based on the instruction of the Lipofectamine 3000 kit (L3000015, Invitrogen, New York, USA), cells were correspondingly assigned into the si-OIP5-AS1 group (2.5 μg), si-NC group (2.5 μg, negative control of si-OIP5-AS1), pcDNA3.1-OIP5-AS1group (2.5 μg), pcDNA3.1-NC group (2.5 μg, negative control of pcDNA3.1-OIP5-AS1), miR-143-3p agomir group (200 nmol), NC agomir (200 nmol, negative control of miR-143-3p agomir), miR-143-3p antagomir group (200 nmol), NC antagomir (200 nmol, negative control of miR-143-3p antagomir), si-OIP5-AS1+miR-143-3p antagomir group, si-NC+NC antagomir group, si-OIP5-AS1+NC antagomir group, si-NC+NC antagomir group, si-NC+miR-143-3p antagomir group and si-NC+NC antagomir group. Blank group was set as the negative control (exposure to the Lipofectamine 3000 kit without any plasmid). Measurement or detection in each group was performed 48 h after cell transfection.
10
miR-199b-3p and AURKA Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MiR‐199b‐3p and AURKA overexpression or silencing vectors, MiR‐199b‐3p‐mimic (miR‐mimic) and its corresponding negative control NC‐mimic, miR‐199b‐3p‐agomir (miR‐agomir), NC‐agomir, miR‐199b‐3p‐inhibitor (miR‐inhibitor), and NC‐inhibitor were constructed. All vectors were obtained from GenePharma. MiR‐199b‐3p‐mimic, miR‐199b‐3p‐agomir, NC‐agomir, miR‐199b‐3p‐inhibitor, sh‐AURKA, and oe‐AURKA and their negative controls were transfected into the cell line HepG2 with Lipofectamine 2000 Reagent (Invitrogen, USA). Then, the transfected cells were placed 2 days under 37°C culture conditions with 5% CO2.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!