Total RNA was isolated from 100 mg of liver samples with the Jena Bioscience Isolation System (Jena Bioscience, Jena, Germany), following the manufacturer's protocol. Total RNA was quantified with a NanoDrop 2000 (Thermo Scientific, Waltham, MA, USA). Reverse transcription was performed with 1 μg of total RNA using the GoScript Reverse Transcription System (Promega) for real-time quantitative PCR, which was analyzed using qPCR GreenMaster with UNG-clear (Jena Bioscience, Jena, Germany) in a StepOne machine (Applied Biosystems) under the following conditions: 50°C for 2 min, 95°C for 3 min, 40 cycles of 95°C for 45 sec, and 60°C for 45 sec. Oligonucleotides were designed to target type I collagen (COL-I), MMP-13, MMP-9, TGFβ, IL-10, IL-1, and β-actin (as a reference control) (Table 1 ). Relative expression level was normalized with β-actin as reference gene, and differences were determined using the 2−ΔΔCt method.
Jena bioscience isolation system
Manufactured by Jena Biosciences
Sourced in Germany
The Jena Bioscience Isolation System is a laboratory equipment designed for the separation and purification of biological molecules, such as proteins, nucleic acids, and other macromolecules. The system utilizes various chromatographic techniques to achieve efficient separation and isolation of target analytes from complex mixtures.
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Lab products found in correlation
2 protocols using jena bioscience isolation system
1
Liver RNA Isolation and qPCR Analysis
2
Quantitative Expression Analysis of Liver Genes
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Total RNA was isolated from 100 mg of each liver sample using the Jena Bioscience Isolation System (Jena Bioscience, Jena, Germany) following the manufacturer's protocol. Total RNA was quantified using a NanoDrop-2000 (Thermo Fisher Scientific, Waltham, MA, USA). Reverse transcription was performed with 1 μg of total RNA using the GoScript Reverse Transcription System (Promega) for real-time quantitative PCR (qPCR), which was performed with the qPCR GreenMaster with UNG-clear (Jena Bioscience, Jena. Germany) in a StepOne machine (Applied Biosystems) with the following conditions: 50°C for 2 min, 95°C for 3 min, 40 cycles of 95°C for 45 s and 60°C for 45 s. Oligonucleotides were designed to target COLL-I, MMP-13, matrix metalloproteinase-9 (MMP-9), transforming growth factor beta (TGF-β), interleukin 6 (IL-6), tissue inhibitor of metalloproteinase-1 (TIMP-1), TIMP-2, V1aR and β-actin (Table 1). Relative expression was normalized to βactin as a reference gene and differences were determined using the 2 -ΔΔCt method.
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