Total RNA from tissue samples or cells was extracted by using Trizol reagent (Invitrogen, CA). RNA was first reversely transcribed into cDNA by using RT reagent Kit (TOYOBO, Japan). Then the cDNA was subjected to RT-PCR with an SYBR Green RT-PCR Master Mix kit (TOYOBO, Japan) in an ABI PRISM 7500 system (Applied Biosystems, USA) by using the miR-126 primers set and U6 primers set (Ribobio, China). The primer sequence of miR-126 is 5′- CATTATTACTTTTGGTACGCGAAA-3′. The relative levels of miR-126 transcripts were normalized to the control U6 mRNA, and the primer sequence was 5′-TCGTGAAGCGTTCCATATTTTTAA-3′. All samples were normalized to internal controls, and the relative expression level was calculated through the 2−ΔΔCt analysis method. Experiments were performed in triplicate samples. Relative gene expression was quantified using the GraphPad Prism 4.0 software (GraphPad Software, San Diego, CA, USA) and expressed as a percentage of the control.
U6 primers set
Manufactured by RiboBio
Sourced in China
The U6 primers set is a laboratory product designed for the detection and analysis of the U6 small nuclear RNA (snRNA). The U6 snRNA is a critical component of the spliceosome, a complex responsible for the removal of introns from pre-mRNA. The U6 primers set provides the necessary tools for researchers to study the expression and function of U6 snRNA in various experimental settings.
Automatically generated - may contain errors
Lab products found in correlation
Rt reagent kit, by Toyobo (2 mentions)
Abi prism 7500 system, by Thermo Fisher Scientific (2 mentions)
Sybr green rt pcr master mix kit, by Toyobo (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Prism 4, by GraphPad (1 mentions)
Mirvana paris kit, by Thermo Fisher Scientific (1 mentions)
Sybr green real time pcr master mix kit, by Toyobo (1 mentions)
2 protocols using u6 primers set
1
Quantification of miR-126 Expression in Tissue and Cell Samples
2
Real-time PCR Quantification of miRNA and mRNA
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Total RNA from the frozen tissues or cultured cells was isolated with mirVanaTM PARISTM Kit (Ambion, USA) according to the manufacturer’s instructions. RNA was first reversely transcribed into cDNA by using RT reagent Kit (TOYOBO, Japan). Then the cDNA was subjected to real-time PCR with an SYBRⓇ Green Realtime PCR Master Mix kit (TOYOBO, Japan) in an ABI PRISM 7500 system (Applied Biosystems, USA) by using the miR-29a primers set and U6 primers set (Ribobio, China). Human VEGF-A primers for RT-PCR were produced by the Sangon Inc., Shanghai, China. The sequence of VEGFA reverse primer is (34) (link) : 5’-ATGATTCTGCCCTCCTCCTT-3’; and the forward primer is: 5’-CCTTGCTGCTCTACC TCCAC-3’ (74 bp). The relative quantification of RNA expression was calculated using the 2-ΔΔCt method (35) (link).
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