RNA interference was conducted using synthetic siRNA duplexes. Two synthetic siRNA duplexes (si-H19) corresponding to the H19 mRNA sequences, 5′-CCCACAACAUGAAAGAAAU-3′ (forward) and 5′-GCUAGAGGAACCAGACCUU-3′ (reverse), were used to inhibit H19 RNA expression. si-H19 and si-negative control (NC) were purchased from Guangzhou RiboBio Co., Ltd. (Guangzhou, China). Cells were cultured in 6-well plates and maintained until they reached ~60% confluence, prior to transfection with siRNA duplexes (25 nM) using Lipofectamine® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocol. Transfection efficiency to assess the inhibition of H19 after 48 h of transfection in Y79 cells was performed using RT-qPCR as previously outlined.
Si h19
Manufactured by RiboBio
Sourced in China
Si-H19 is a laboratory instrument used for RNA detection and quantification. It utilizes the H19 gene as a target for analysis. The core function of Si-H19 is to provide accurate and reliable measurements of H19 gene expression levels in samples.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Mir 130a 3p mimic, by RiboBio (1 mentions)
Ribofecttm, by RiboBio (1 mentions)
Pcdna3.1 h19, by GenePharma (1 mentions)
Lipofectamine 3000 transfection reagent, by Thermo Fisher Scientific (1 mentions)
Mir 17 5p inhibitor, by GenePharma (1 mentions)
Mir 130a 3p mimic, by GenePharma (1 mentions)
Mir 130a 3p inhibitor, by GenePharma (1 mentions)
Pcdna3, by Thermo Fisher Scientific (1 mentions)
Mir 17 5p mimic, by GenePharma (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Si nc, by RiboBio (1 mentions)
Mir nc, by GenePharma (1 mentions)
5 protocols using si h19
1
Silencing H19 RNA Expression
2
Transfection of NSCLC Cells with miR-130a-3p and siRNAs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NSCLC cells were plated at 50% confluence in 35-mm Petri dishes. RiboFECTTM (RiboBio, China) was used as a transfection reagent. The concentration of both the miR-130a-3p mimic and si-H19 (RiboBio, China) was 50 nM, and that of si-WNK3 (RiboBio, China) was 100 nM. After a further 24 h, cells were harvested. The transfection efficiency of the siRNA and mimic was measured relative to the expression of a housekeeping gene by qRT–PCR.
lncRNAH19: siRNA: CCTCTAGCTTGGAAATGAA.
WNK3: siRNA: GACCGACAGTTGTTTCACA.
lncRNAH19: siRNA: CCTCTAGCTTGGAAATGAA.
WNK3: siRNA: GACCGACAGTTGTTTCACA.
3
Plasmid and RNA Modulation of H19
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pcDNA3.1-H19, a plasmid for H19 overexpression, was sourced from the GenePharma (Shanghai, China). si-H19, a small interfering RNA that reduces H19 expression, was obtained from RiboBio. miR-675-5p inhibitors and mimics, which can reduce or overexpress miR-675-5p, were obtained from RiboBio.
4
Transfection of H19 and TDG in SRA01/04 cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SRA01/04 cells were cultured in 6‐well plates to 60%‐70% confluence prior to transfection. Si‐H19, Si‐H19 NC, si‐TDG, si‐TDG NC, miRNA‐29a mimic and mimic control were purchased from RiboBio (Guangzhou, China). H19 cDNA was amplified and cloned into pcDNA vector (Hanyinbt, Shanghai, China). The siRNA or vector was transfected into cells with lipofectamine 3000 transfection reagent (Invitrogen, USA) according to the manufacturer's protocol.
5
Transfection of Plasmids and Oligonucleotides in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The pcDNA3‐H19 and pcDNA3 were provided by Invitrogen (Carlsbad, CA, USA), and si‐H19 and si‐NC were synthesized by Ribobio (Guangzhou, China) based on siDirect software (https://maidesigner.invitrogen.com/rnaexpress ; Table S1 ). Moreover, the miR‐130a‐3p mimic, miR‐130a‐3p inhibitor, miR‐17‐5p mimic, miR‐17‐5p inhibitor, and miR‐NC were purchased from Genepharma (Shanghai China). The transfections of plasmid and oligonucleotide were performed utilizing Lipofectamine 2000 reagent (Life Technologies). After transfection for 48‐72 hours, the cells were harvested for further experiments.28
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!