Total RNA were isolated from fresh‐frozen tissues and cultured cells using TransZol reagents (TransGen Biotech). Reverse transcription was carried out using a Transcriptor First Strand cDNA Synthesis Kit (Roche, Indianapolis, IN, USA) according to the manufacturer's instructions. The divergent primers for circADAMTS13, circDPF3, and circCASP8AP2 were designed to determine the abundance of circRNA. Bulge‐Loop™ hsa‐miR‐484 qRT‐PCR Primer Set (RiboBio, Guangzhou, China) and U6 snRNA qPCR Primer Set (RiboBio) were used for amplification of miR‐484 and U6, respectively. The other primers are listed in Table S1 . For qRT‐PCR analysis, aliquots of double‐stranded cDNA were amplified using SYBR Premix Ex Taq II (Takara, Dalian, China). The relative gene expression was normalized to the geometric mean of the housekeeping gene 18SrRNA and then calculated according to the Livak method (∆∆Ct; Livak and Schmittgen, 2001 ). The experiments were independently repeated three times.
U6 snrna qpcr primer set
Manufactured by RiboBio
Sourced in China
The U6 snRNA qPCR Primer Set is a laboratory equipment product designed for the quantitative real-time PCR (qPCR) analysis of U6 small nuclear RNA (snRNA) expression. U6 snRNA is a widely used internal control or reference gene for gene expression studies. The primer set provides the necessary primers for the specific amplification of U6 snRNA sequences during qPCR experiments.
Automatically generated - may contain errors
Lab products found in correlation
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
Transzol reagent, by Transgene (1 mentions)
Sybr premix ex taq 2, by Takara Bio (1 mentions)
Transcriptor first strand cdna synthesis kit, by Roche (1 mentions)
Bulge loop mirna qrt pcr primer set, by RiboBio (1 mentions)
Bulge loop mirna qrt pcr starter kit, by RiboBio (1 mentions)
Total rna extraction kit, by Generay (1 mentions)
Iq sybr green supermix, by Bio-Rad (1 mentions)
2 protocols using u6 snrna qpcr primer set
1
Circularized RNA Quantification Protocol
2
Quantification of miRNA and mRNA Expression
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Total RNA was extracted from the cells using a Total RNA Extraction Kit (Generay Biotech, Shanghai, China), then RNA was reverse-transcribed into cDNA using a RevertAid First Strand cDNA synthesis Kit (Thermo Scientific Fisher, Waltham, MA, UK) and the thermocycling program used was as follows: 37℃ for 60 mins and 85℃ for 5mins. Amplifications were performed in an iCycler using iQ SYBR Green supermix (Bio-Rad). Gapdh was amplified on the same plates and used to normalize the data. Each sample was prepared in triplicate and each experiment was repeated at least three times. The relative abundance of each gene was quantified using the 2 -ΔΔCt method. The PCR primers used are listed in Table S1. Total RNA for miR-3605-5p, miR-6082-3p, and U6 detection was extracted with a Total RNA Extraction Kit. Reverse transcription and PCR was performed using a BulgeLoopTM miRNA qRT-PCR Starter Kit, a Bulge-Loop™ miRNA qRT-PCR Primer Set, and a U6 snRNA qPCR Primer Set (RiboBio, Guangzhou, China) according to the manufacturer's instructions. miRNA expression was quantified using the 2 -ΔΔCt method and U6 was used as an internal control.
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