Rnaprep pure tissue kit

Manufactured by Tiangen Biotech

Sourced in China, United States

The RNAprep Pure Tissue Kit is a product designed for the efficient isolation of high-quality total RNA from various tissue samples. It utilizes a silica-based membrane technology to ensure the selective binding and purification of RNA molecules.

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Close spelling variants of this product

RNAprep Pure Kit (104 citations) RNAprep Tissue Kit (4 citations) RNAprep Pure kit (For Tissue) (4 citations) RNAprep Pure Animal Tissue Total RNA Extraction Kit (2 citations) RNAprep pure animal tissue kit (2 citations)

230 protocols using rnaprep pure tissue kit

Quantifying mRNA and miRNA Levels from Tissue Samples

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For measurement of mRNA levels, total RNA from tissue samples was isolated using RNAprep Pure Tissue kit (Tiangen, Beijing, China) according to the manufacturer’s instructions. Random-primed cDNAs were generated by reverse transcription of total RNA with TIANScript M-MLV (Tiangen). A real-time PCR analysis was performed with the ABI Prism 7000 Sequence Detection system (Applied Biosystems) using SYBR® Premix-Ex TagTM (Takara, Dalian, Chnia). GAPDH was used for an internal control. Primer sequences for these mRNAs were listed in Table S2.
Mature miRNAs from tissue samples, serum and exosomes were extracted using miRcute miRNA Isolation kit (Tiangen). Pre-miRNAs from tissue samples were extracted using RNAprep Pure Tissue kit (Tiangen). poly(A) modification and first-strand cDNA synthesis were performed with miRcute miRNA first-strand cDNA synthesis kit (Tiangen). qRT-PCR analysis for mature miRNA was conducted using miRcute miRNA qRCR detection kit (Tiangen). For measurement of miRNA or pre-miRNA levels in the tissue samples,18S rRNA was used as an internal control. The miRNA levels in serum and exosomes were determined using absolute quantitative real-time RT-PCR described previously^{40 (link)}. Primer sequences for these miRNAs were listed in Table S2.

Zou T., Zhu M., Ma Y.C., Xiao F., Yu X., Xu L., Ma L.Q., Yang J, & Dong J.Z. (2018). MicroRNA-410-5p exacerbates high-fat diet-induced cardiac remodeling in mice in an endocrine fashion. Scientific Reports, 8, 8780.

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Quantification of Gene Expression in Jejunum Mucosa

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Total RNA was isolated from jejunum mucosa using RNAprep Pure Tissue Kits (Tiangen Biotech Co., Ltd., Beijing, China). OD260/OD280 analyses and agarose gel (1.2%) electrophoresis were performed to evaluate RNA purity and integrity. Reverse transcription was performed using PrimeScript RT Mix Kits (TaKaRa, Kusatsu, Japan) as per manufacturer's instructions. RT-PCR analyses were conducted using a CFX96 PCR System (Bio-Rad, Hercules, CA) using TB Green Premix Ex Taq kits (TaKaRa). Relative mRNA expression levels were calculated using the 2^−ΔΔCt method. Primers are listed (Supplementary Table S2).

Liu H., Meng H., Du M., Lv H., Wang Y, & Zhang K. (2024). Chlorogenic acid ameliorates intestinal inflammation by inhibiting NF-κB and endoplasmic reticulum stress in lipopolysaccharide-challenged broilers. Poultry Science, 103(5), 103586.

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Quantitative RT-PCR Analysis of Osteogenic Gene Expression

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Total RNA was extracted and detected using RNAprep Pure tissue kits (TIANGEN Biotech Co., Ltd, Beijing, China) and a NanoDrop ND-1000 Spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA), respectively. Meanwhile, cDNA was synthesized according to the instructions of the Takara PrimeScript RT reagent kit gDNA Eraser (TaKaRa Biotech, Kyoto, Japan) in accordance with the manufacturer's instructions. Then, qRT-PCR was performed on an ABI 7500 quantitative polymerase chain reaction system (Life Technologies Inc., Carlsbad, CA, USA). In detection of ANCR expression (forward, 5′-GCCACTATGTAGCGGGTTTC-3′; reverse, 5′-CCTGCGCTAAGAACTGAGG-3′), Runx2 expression (forward, 5′-CGGGTCTCCTTCCAGGAT-3′; reverse, 5′-GGGAACTGCTGTGGCTTC-3′), Osterix expression (forward, 5′-GAAGCGACCACTTGAGCACAT-3′; reverse, 5′-TGTCCAAACTCATCAATGTATCT-3′), and EXH2 expression (forward, 5′-TTGTTGGCGGAAGCGTGTAAAATC-3′; reverse, 5′-TCCCTAGTCCCGCGCAATGAGC-3′), the PCR program included 95℃ for 10 min, 40 cycles of 95℃ for 10 s, 60℃ for 20 s, and 72℃ for 34 s. GAPDH was used as an internal control (forward: 5′-CGAGCCACATCGCTCAGACA-3′; reverse: 5′-GTGGTGAAGACGCCAGTGGA-3′). The experiment was repeated three times. The relative expression of target genes was calculated using the 2^−ΔΔCt method.15 (link)

Cai N., Li C, & Wang F. (2019). Silencing of LncRNA-ANCR Promotes the Osteogenesis of Osteoblast Cells in Postmenopausal Osteoporosis via Targeting EZH2 and RUNX2. Yonsei Medical Journal, 60(8), 751-759.

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Quantifying SYPL2 Expression in CRC

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Total RNA extracted from cells using RNAprep Pure Tissue kits (Tiangen, Beijing, China) was reverse-transcribed to complementary DNA using the FastKing gDNA Dispelling RT SuperMix for quantitative polymerase chain reaction (qPCR) (Tiangen). The samples were subjected to quantitative real-time PCR (qRT-PCR) using 2 × SYBR Green qPCR Master Mix (Tiangen) and primers specific for SYPL2 (forward, 5’-CGCTGGTGGACTTCTGTG-3’; reverse, 5’-GCTGGATGGTCGTGTGG-3’) and GAPDH (forward, 5’-AAGGTCGGAGTCAACGGA-3’; reverse, 5’-TTAAAAGCAGCCCTGGTGA-3’), with all gene primers obtained from Aoke Dingsheng Biotechnology (Beijing, China). Thermal cycling conditions consisted of an initial denaturation at 95 ºC for 15 s, followed by 40 cycles of denaturation at 95 ºC for 15 s, annealing at 55 °C for 30 s, and extension at 72 ºC for 30 s. Relative messenger RNA expression levels were calculated using the 2^-∆∆CT method, with the average level of SYPL2 expression in the 20 normal colorectal tissues defined as the reference for normalization and comparison with the 20 CRC tissues.

Zhao Z.X., Liu Q.L., Yuan Y, & Wang F.S. (2022). Synaptophysin-like 2 expression correlates with lymph node metastasis and poor prognosis in colorectal cancer patients. World Journal of Gastrointestinal Oncology, 14(11), 2122-2137.

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RNA Extraction and Real-Time PCR Analysis

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Total RNA was extracted using an RNAprep Pure tissue kit or RNAprep Pure cell kit (Tiangen, Beijing, China). The total RNA (2 μg) was then reverse transcribed to cDNA using the FastKing gDNA Dispelling RT SuperMix (Tiangen, China). Quantitative real-time PCR was performed in triplicate using the SYBR Green qPCR SuperMix (Transgen Biotech, Beijing, China) on an ABI Q6 instrument (Applied Biosystems, Waltham, MA, USA) according to the manufacturer’s instructions. Gene expression was normalized against the murine or human glyceraldehyde-3-phosphate dehydrogenase (GAPDH) housekeeping gene. The primers used are listed in Supplementary Table S2.

Hong F., Pan S., Xu P., Xue T., Wang J., Guo Y., Jia L., Qiao X., Li L, & Zhai Y. (2020). Melatonin Orchestrates Lipid Homeostasis through the Hepatointestinal Circadian Clock and Microbiota during Constant Light Exposure. Cells, 9(2), 489.

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RNA Extraction from Bacterial Coaggregates

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Total RNA of the coaggregated pellets and the monocultures of F. nucleatum subsp. polymorphum and S. gordonii DL1 was extracted using the hot phenol method and RNAprep Pure Tissue Kit according to the manufacturer’s instructions (Tiangen Biotech, Beijing, China). The extracted RNA was dissolved in 30-100 μl RNase-free water. RNA was quantified using NanoDrop-2000 (NanoDrop Technologies, DE, USA), and the integrity of RNA was determined using 2100 Bioanalyser (Agilent, CA, USA).

Liu T., Yang R., Zhou J., Lu X., Yuan Z., Wei X, & Guo L. (2022). Interactions Between Streptococcus gordonii and Fusobacterium nucleatum Altered Bacterial Transcriptional Profiling and Attenuated the Immune Responses of Macrophages. Frontiers in Cellular and Infection Microbiology, 11, 783323.

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RT-qPCR Gene Expression Analysis

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Total RNA was extracted using an RNAprep Pure Tissue Kit (Tiangen, Beijing, China), and it was reverse-transcribed to generate cDNA using PrimeScript RT Master Mix (TaKaRa, Dalian, China) according to the manufacturer’s instructions. RT-qPCR was performed on an ABI 7500 system (Applied Biosystems, Shanghai, China) using FastStart Universal SYBR Green Master Mix (ROX; Roche Diagnostics). The primers used in RT-qPCR are listed in Table 1. Gene expression levels were calculated using the 2^−∆∆CT method and were normalized to β-actin mRNA expression.

Ying S., Qin J., Dai Z., An H., Zhu H., Chen R., Yang X., Wu W, & Shi Z. (2020). Effects of LPS on the Secretion of Gonadotrophin Hormones and Expression of Genes in the Hypothalamus-Pituitary-Ovary (HPG) Axis in Laying Yangzhou Geese. Animals : an Open Access Journal from MDPI, 10(12), 2259.

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Quantitative Real-Time RT-PCR Analysis

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Total RNA from tissue samples and cells were extracted with RNAprep pure tissue kit (Tiangen Biotech Co., LTD., Beijing, China) and Rneasy Mini Kit (Qiagen, Germantown, MD, USA) respectively. The mRNA of DNMT1, miR-124-3p and BCAT1 were amplified by quantitative real-time RT-PCR. It was carried out using SuperScriptIII Platinum SYBR Green One-Step qRT-PCR kit (Invitrogen) by an ABI 7500 Fast real-time PCR instrument (Applied Biosystems, Foster City, CA, USA). It was carried out with the following procedures: 50 °C for 3 min, 95 °C for 5 min, followed by 35 cycles of 95 °C for 15 s, 60 °C for 30 s. The specific primers used in quantitative real-time RT-PCR were displayed as follows: DNMT1: forward, 5′-CCTAGCCCCAGGATTACAAGG-3′; reverse, 5′-ACTCATCCGATTTGGCTCTTTC-3′; BCAT1: forward: 5′- GAGCCTGGAAAGGTGGAACTG-3′; reverse, 5′- GCTGACACCCATTATCTACTGCT-3′; GAPDH: forward, 5′-TGTTCGTCATGGGTGTGAAC-3′; reverse, 5′-ATGGCATGGACTGTGGTCAT-3′; hsa-miR-124-3p: forward, 5′-ACACTC CAGCTGGGTAAGGCACGCGGTGAA-3′; reverse, 5′- CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGAGGGCATTCAC-3; U6: forward, 5′-CTCGCTTCGGCAGCACA-3′; reverse, 5′-AACGCTTCACGAATTTGCGT-3′. The data were analyzed according to the comparative Ct method.

Zeng B., Zhang X., Zhao J., Wei Z., Zhu H., Fu M., Zou D., Feng Y., Luo H, & Lei Y. (2019). The role of DNMT1/hsa-miR-124-3p/BCAT1 pathway in regulating growth and invasion of esophageal squamous cell carcinoma. BMC Cancer, 19, 609.

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Total RNA Extraction and cDNA Synthesis

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The total RNA was extracted from animal tissues (intestine, liver, gill, heart, kidney, brain, muscle, spleen, skin, and stomach) using total RNA Extraction Kits (RNAprep pure Tissue Kit, Tiangen Biotech Co., Ltd., Beijing, China), and the quality and concentration of RNA were detected by 1% agarose gel electrophoresis and Nanodrop 2000 (Thermo Fisher Scientific, Waltham, MA). The RNAs that met the quality criteria (28S:18S = 2:1) were used to synthesize cDNA according to the steps of the reverse transcription kit (TranScript One-Step gDNA Removal and cDNA Synthesis SuperMix, TransGen Biotech Co., Ltd., Beijing, China), and the samples were stored at −20 °C for analysis.

Wang X., Zhao T, & Ma A. (2022). Genetic Mechanism of Tissue-Specific Expression of PPAR Genes in Turbot (Scophthalmus maximus) at Different Temperatures. International Journal of Molecular Sciences, 23(20), 12205.

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RNA Extraction from Mammary Tissue

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For RNA extraction, the mammary tissue was treated with TRIzol reagent (Invitrogen, Carlsbad, CA, USA), and the total RNA was then extracted using the RNAprep Pure Tissue Kit (Tiangen, RNAprep Pure Tissue Kit, Beijing, China). Briefly, frozen mammary tissue (10–20 mg) was combined with 300 μL lytic solution RL and was thoroughly ground with a grinding pestle. Then, 590 μL RNase-free ddH2O and 10 μL Proteinase K were added to the homogenate and mixed at 56 °C for 10–20 min. After dissolving the total RNA with appropriate diethylpyrocarbonate (DEPC), the quantity and quality of RNA were measured using a spectrophotometer (NanoDrop^® ND-1000, Thermo Scientific, DE, Waltham, MA, USA). The quantity of total RNA was greater than 400 ng μL⁻¹, and the 260/280 requirement was 1.9~2.0.

Liang Y., Gao Q., Wang H., Guo M., Arbab A.A., Nazar M., Li M., Yang Z., Karrow N.A, & Mao Y. (2022). Identification and Characterization of Circular RNAs in Mammary Tissue from Holstein Cows at Early Lactation and Non-Lactation. Biomolecules, 12(3), 478.

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