Genomic DNA was extracted using salting out method. PCR products with altered gel mobility were sequenced. All PCR products were sequenced with Rotor Gene PCR Analyzer (Qiagen, Germany) using the BIG DYE terminator V1.1 cycle sequencing kit.
Rotor gene pcr analyzer
Manufactured by Qiagen
Sourced in Germany
The Rotor-Gene PCR Analyzer is a real-time PCR instrument designed for nucleic acid analysis. It features a rotary design that allows for precise temperature control and uniform heating of samples. The Rotor-Gene PCR Analyzer is capable of performing real-time PCR, quantitative PCR, and melt curve analysis.
Automatically generated - may contain errors
2 protocols using rotor gene pcr analyzer
1
DNA Extraction and Sequencing Protocol
2
Quantitative Analysis of NF-κB Gene Expression
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After 14 days of incubation in Osteogenic media, extraction of a total of mRNA was achieved using the RNeasy Mini Kit (Qiagen, Hildesheim, Germany). in accordance with the manufacturer's instructions, using the Quantitect RT Kit (Qiagen, Hilden, Germany) Table 1 . “Quantification” of NF-k gene expression was amplified from total RNA extracts using Quantitect primer assay - primer assays, cat no: 249900; [Hs_NF-k, ID: QT00396823]. Primordial Test. Quantitect Syber green Master mix served to amplify the genes (Qiagen, Hilden, Germany). Similarly in earlier research, the -actin (Hs ACTB) primer assay (ID: QT000954231) was utilized as a housekeeping gene (Abdelgawad et al., 2021 (link)). The five-plex Rotor-Gene PCR Analyzer was used to evaluate all samples (Qiagen, Germany). Gene expression levels were analyzed using the ΔCt technique, with B-actin serving as a housekeeper gene for normalization. The tests were repeated in triplicate.
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