Total RNA from ASCs was isolated with the miRNeasy Mini Kit (217004, Qiagen, Germany), in accordance with the manufacturer’s instructions. RNA was transcribed to cDNA using the HiScript® II RT kit (R233, Vazyme, Nanjing, China) and Mir-X™ miRNA First-Strand Synthesis kit (RR638315, TaKaRa) separately. Quantitative polymerase chain reaction (qPCR) was performed using the HiScript® II One Step qRT-PCR SYBR Green Kit (Q221, Vazyme). miR-143-3p RNA was reverse transcribed using a particular RT primer (Tsingke, China). The U6 and GAPDH genes were used as references for the amplification of miRNAs and other genes. Expression levels of target genes were assessed using the 2−ΔΔCt method. The primer sequences are described in Additional file 1 : Table S2. The RNA-seq data underlying this article have been uploaded to the Gene Expression Omnibus (http://www.ncbi.nlm.nih.gov/geo ; GSE174502).
Hiscript 2 rt kit
Manufactured by Vazyme
Sourced in China
The HiScript® II RT kit is a reverse transcription kit for the synthesis of complementary DNA (cDNA) from RNA templates. The kit contains all the necessary components, including reverse transcriptase enzyme, reaction buffer, and RNase inhibitor, to facilitate the conversion of RNA to cDNA.
Automatically generated - may contain errors
Lab products found in correlation
Mir x mirna first strand synthesis kit, by Takara Bio (2 mentions)
Mirneasy mini kit, by Qiagen (2 mentions)
Hiscript 2 one step qrt pcr sybr green kit, by Vazyme (1 mentions)
Hiscript 2 one step rt pcr sybr green kit, by Vazyme (1 mentions)
Steponeplus platform, by Thermo Fisher Scientific (1 mentions)
Cytoplasmic nuclear rna purification kit, by Norgen Biotek (1 mentions)
2 protocols using hiscript 2 rt kit
1
Quantification of miR-143-3p in ASCs
2
Isolation and Detection of RNAs
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Total RNAs were isolated using the miRNeasy Mini Kit (Qiagen, Germany). miRNAs were transcribed into cDNAs using the Mir-X miRNA First Strand Synthesis kit (#RR638315, TaKaRa), while mRNAs and lncRNAs were transcribed into cDNAs using the HiScript II RT kit (Vazyme, Nanjing, China). Real-time PCR was detected under the StepOnePlus platform (Applied Biosystems) using the HiScript II One Step RT-PCR SYBR Green Kit (Vazyme). The relative expression levels of the targeted genes were normalized to β-actin, GAPDH, and U6, and then calculated using the 2−ΔΔCt method. The cytoplasmic and nuclear RNAs were isolated using the Cytoplasmic & Nuclear RNA Purification Kit (Norgen Biotek Corp). The RNAs were then transcribed into cDNAs and detected using the above methods. GADPH and U3 were used as cytoplasm-located and nucleus-located markers separately. The primers used in the PCR were described in Supplementary Tables S3 and S4 .
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