Tripure reagent

Manufactured by Takara Bio

Sourced in China

TRIpure Reagent is a ready-to-use solution for the isolation of total RNA from various biological samples. It is a mono-phasic solution of phenol, guanidine isothiocyanate, and other proprietary components that facilitate the effective lysis of cells and tissues, and the subsequent isolation of intact RNA.

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TRIpure Reagent kit (12 citations)

2 protocols using tripure reagent

RNA Extraction and Analysis from Kidney, Plasma, and Urine

Cited 1 time

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Kidney tissues were quickly ground in liquid nitrogen, and total RNA was extracted using the TRIpure Reagent (Takara, Dalian, China) according to the manufacturer’s instructions. Plasma RNA was extracted with a BIOG cfRNA Easy Kit (BIOG, Changzhou, China). Urine from clinical subjects was centrifuged at 3,200 × g for 20 min, and the sediment was washed with PBS before centrifugation at 12,000 × g for 5 min. Then, the total RNA was extracted and qualified by the measurement of A260/A280 ratio, which indicated a high purity of the extracted RNA ranged from 1.8 to 2.0. RNA samples were reverse transcribed and amplified as previously described.⁶³ (link)
β-actin and/or U6 were used as internal controls, and the relative expression of different genes was calculated following the 2^–ΔΔCT method or following a log transformation after the 2^–ΔCT method. The primer sequences are listed in Table S5.

Wang Z., Chang Y., Liu Y., Liu B., Zhen J., Li X., Lin J., Yu Q., Lv Z, & Wang R. (2022). Inhibition of the lncRNA MIAT prevents podocyte injury and mitotic catastrophe in diabetic nephropathy. Molecular Therapy. Nucleic Acids, 28, 136-153.

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Comprehensive RNA Extraction and Quantification

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TriPure Reagent (Takara Bio Inc., Dalian, China) was used for isolating total RNA from ventricular tissue. The tissue was directly lysed by mixing with 1 ml of TriPure Reagent and homogenized using a homogenizer. Then 200 μl of chloroform was added to the homogenized sample and incubated for 25 min at room temperature. Subsequently, RNA was precipitated by mixing with isopropyl alcohol. An ultraviolet spectrophotometer was used to measure the absorbance of RNA at 260 and 280 nm, and the concentration of RNA was determined. Then mRNA was isolated from total RNA using Oligo (dT), and reverse transcribed into first‐strand complement DNA (cDNA) and amplified using an SYBR GREEN Master Mix cDNA synthesis kit. The reaction system included 1 μl of cDNA, 10 μl of 2 × SYBR GREEN Master Mix, and 0.5 μl of each primer. The PCR condition was as follows: preincubation at 94°C for 2 min, followed by 40 cycles of denaturation at 94°C for 15 s, and annealing/extension at 60°C for 30 s using Exicycler™ 96 detection system. The primers are listed in Table 1.

Liang C., Zhang T., Shi X., Jia L., Wang Y, & Yan C. (2021). Modified Renshen Yangrong decoction enhances angiogenesis in ischemic stroke through promotion of MicroRNA‐210 expression by regulating the HIF/VEGF/Notch signaling pathway. Brain and Behavior, 11(8), e2295.

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