An RNA isolation system (Sigma-Aldrich, St. Louis, MO, USA) was used to extract total RNA and a Prime Script RT reagent kit with a gDNA removal kit (Takara Bio, Otsu, Japan) was used in obtaining cDNA. A 20 µL total reaction system containing 10 uL of SYBR (Takara Bio, Otsu, Japan), 0.5 µM forward primer, 0.5 µM reverse primer, 5 ng/µL cDNA, and DEPC water was used. A qRT-PCR machine was set at 95 °C for 2 min followed by 40 cycles of 95 °C for 15 s and 60 °C for 30 s. Data were analyzed using comparative 2−△△Ct with normalization to the Gapdh housekeeping gene. The primers were synthesized in Kumei, Changchun, China (Table 1 ).
Rna isolation system
Manufactured by Merck Group
Sourced in United States
The RNA isolation system is a laboratory equipment designed to extract and purify RNA from various biological samples. It utilizes a series of processing steps to separate and concentrate the RNA component from the sample, allowing for further analysis and applications.
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Lab products found in correlation
2 protocols using rna isolation system
1
Quantitative Transcript Analysis of Gene Expression
2
Quantitative Real-Time PCR Protocol
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Total RNA was obtained using an RNA isolation system (Sigma-Aldrich), and then the previously mentioned extracted RNA was reverse transcribed using a PrimeScript RT kit with gDNA removal function (Takara Bio). For cDNA, the experimental procedure was performed in strict accordance with the manufacturer's instructions. The total qRT-PCR reaction system was 20 µL, containing 10 µL SYBR (Takara Bio), 0.5 µM for the forward primer, 0.5 µM for the reverse primer, 5 ng/µL for the cDNA, and 20 µL supplemented with sterile ultrapure water. The reaction conditions were as follows: 95°C for 2 min, 95°C for 15 s, and 60°C for 30 s for 40 cycles. The 2 -ΔΔCt method was used to calculate the relative expression of genes. The primers were synthesized from KuMei, Changchun, China (Table 1).
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