RNA was extracted using an RNAeasy™ Animal RNA isolation kit with a spin column (Beyotime, #R0026), and miRNA was extracted using a SanPrep column miRNA extraction kit (Sangon Biotech, #B518811), according to the manufacturer’s instructions. RNA reverse transcription, cDNA verification, preparation of a real-time PCR system, and determination of the reaction conditions were carried out as described previously. β-actin and U6 were the reference genes. The primers are listed in Table 3
Sanprep column mirna extraction kit
Manufactured by Sangon
The SanPrep column miRNA extraction kit is a tool designed to isolate and purify microRNA (miRNA) from various biological samples. It utilizes a silica-based column chromatography technique to selectively capture and concentrate miRNA molecules. The kit provides a straightforward protocol for efficient miRNA extraction, enabling researchers to obtain high-quality miRNA samples for downstream analysis and applications.
Automatically generated - may contain errors
Lab products found in correlation
Rnaeasy animal rna isolation kit, by Beyotime (1 mentions)
Mirna qpcr kit, by Sangon (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Transcriptor first strand cdna synthesis kit, by Roche (1 mentions)
Faststart universal sybr green master rox, by Roche (1 mentions)
Mirna first strand cdna synthesis, by Sangon (1 mentions)
2 protocols using sanprep column mirna extraction kit
1
Isolation and Analysis of miRNA and mRNA
2
Comprehensive RNA extraction and qRT-PCR analysis
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Total RNA from cells and tissues were extracted using Trizol reagent (Invitrogen, USA) following the manufacturer’s instructions. For lncRNAs and mRNAs, Transcriptor First Strand cDNA synthesis Kit (Roche, 0489703000) was applied for reverse transcriptions. qRT-PCR was performed using FastStart Universal SYBR Green Master (Rox) (Roche, 4913850001) on the instrument of ABI 7500 (Thermo, USA). For miRNAs, SanPrep Column miRNA Extraction Kit(Sangon Biotech, B518811), miRNA First Strand cDNA Synthesis (Sangon Biotech, B532453) and miRNA qPCR Kit (Sangon Biotech, B532461) were adopted. Relative gene expressions were determined using the 2–ΔΔCT method. The primers were described in Table 1 .
Primer sequences used for qRT-PCR
| Gene | Primer | Sequence (5′-3′) |
|---|---|---|
| GADPH | Forward | GCATCCTGGGCTACACTG |
| Reverse | TGGTCGTTGAGGGCAAT | |
| U6 | Forward | CGCTTCGGCAGCACATATACTAAAATTGGAAC |
| Reverse | GCTTCACGAATTTGCGTGTCATCCTTGC | |
| RT Primer | AAAATATGGAACGCTTCACG | |
| lncRNA XIST | Forward | AGCAGGTCAGGCAGAGGAAGTC |
| Reverse | CCCGATACAACAATCACGCAAAGC | |
| TNFSF10 | Forward | TTACCAACGAGCTGAAGCAGATGC |
| Reverse | GCTGACGGAGTTGCCACTTGAC | |
| miR-129-5p | Forward | TGCGCTTTTTGCGGTCTGG |
| Reverse | AGTGCAGGGTCCGAGGTATT | |
| RT Primer | GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACGCAAGC |
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