Forty-eight of the original 78 samples that had enough tissue were used for total RNA isolation. Total RNA was isolated using RNeasy Mini Kit (QiaGen, MD, USA), and converted to cDNA using the High Capacity cDNA Reverse Transcription Kits (Applied Biosystems®, Life Technologies, CA, USA). GSTZ1 gene expression level was measured by Taqman Gene Expression Assays (TaqMan gene expression probes; Hs99999901_s1 for 18S, and Hs01041668_m1 for GSTZ1) on a Taqman 7900 Real Time PCR system (Applied Biosystems®, Life Technologies, Carlsbad, CA, USA). GSTZ1 gene expression levels were normalized to the reference gene 18S (ribosomal RNA). The 2−ΔCt method was used to assess relative gene expression differences between samples carrying different GSTZ1 haplotypes [16 (link)]. Samples were grouped as those who carried at least one copy of the K allele, and those who carried no copies of the K allele.
Taqman 7900 real time pcr system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Taqman 7900 Real Time PCR system is a laboratory instrument designed for real-time quantitative PCR (qPCR) analysis. It utilizes Taqman chemistry to detect and quantify target DNA sequences in real-time. The system includes optical detection capabilities, temperature control, and data analysis software to facilitate accurate and reliable qPCR experiments.
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2 protocols using taqman 7900 real time pcr system
1
GSTZ1 Gene Expression Quantification
2
Quantifying Gene Expression in Mouse Brain
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High-quality total RNA was isolated from mouse brain tissue using Qiagen RNeasy Isolation Kit (Qiagen). mRNA expression of 4 selected genes (choline acetyltransferase (Chat), acetylcholinesterase (Ache), allograft inflammatory factor 1 (Aif1), the gene coding ionized calcium-binding adapter molecule 1 (Iba1), beta-site APP cleaving enzyme 1 (Bace1)) was quantified in mouse brain tissues using predesigned TaqMan Gene Expression Assays (Mm01221882_m1, Mm00477274_g1, Mm00479862_g1, Mm00478664_m1) from Applied Biosystems using the TaqMan 7900 Real-Time PCR system (Applied Biosystems). Raw data were normalized to Gapdh (Mm99999915) as a housekeeping gene. Relative expression quantification was achieved with the comparative ΔΔCt method.
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