RNA from 50–100 × 103 T cells was purified using Stratagene RNA kit and directly transferred into the RT reagent using the Applied Biosystems Taqman reverse transcriptase reagents. Samples were then subjected to real-time PCR analysis on the Applied Biosystems PRISM 7000 Sequencer Detection System (Applied Biosystems, Foster City, CA, USA) using standard conditions. Genes analyzed were detected with commercially available assays (Applied Biosystems). The relative mRNA abundance was normalized against GAPDH.
Prism 7000 sequencer detection system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PRISM 7000 Sequencer Detection System is a laboratory instrument used for DNA sequencing. It performs automated, real-time detection and analysis of DNA samples. The system is designed to provide reliable and accurate sequencing data for research and diagnostic applications.
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Lab products found in correlation
4 protocols using prism 7000 sequencer detection system
1
Quantitative RT-PCR Analysis of T Cells
2
RNA Purification and Real-Time PCR Analysis
3
Real-Time qPCR Analysis of Gene Expression
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RNA was purified using Stratagene RNA kit and transferred directly into the RT reagent using the Applied Biosystems Taqman reverse transcriptase reagents. Samples were subjected to real-time PCR analysis on an Applied Biosystems PRISM 7000 Sequencer Detection System (Applied Biosystems, Foster City, CA) under standard conditions. Genes analyzed were detected using commercially available assays (Applied Biosystems). For Foxo1, assay ID#: Mm00490671_m1; for Muc5ac, assay ID# Mm01276718_m1; for Irf4, assay ID Mm00516431_m1; for PU.1, assay ID Mm03048233_m1. Relative mRNA abundance was normalized against GAPDH, assay ID Mm99999915_g1 or 18 s, assay ID Mm03928990_g1.
4
Quantitative Analysis of AHI1 Expression
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RNA was isolated using the Qiagen Plus Micro Kit (Qiagen, Venlo, the Netherlands) and converted to cDNA via reverse transcriptase by random hexamers and MuLV transcriptase (Applied Biosystems, Foster City, CA). Samples were subjected to real-time PCR analysis on PRISM 7000 Sequencer Detection System (Applied Biosystems) under standard conditions. The primers used for the human AHI1 were purchased from Applied Biosystems: isoform A forward primer 5′- CCAGCTAATCATGTGGCTAGTGAAACACTG-3′; reverse primer 5′ CCTCAGGGCTTAAAGGAGGGGATGC-3′; isoform B forward primer 5′- TCAGACCGACAGTCACTTTGCTGAA-3′; and reverse primer 5′ TGGTGGGATCCCAGGTCGGCTCAGT-3′. Values are represented as the difference in Ct values normalized to β2-microglobulin for each sample as per the following formula: relative RNA expression = (2−dCt) × 103. Murine Ahi1 was measured using commercially available assays (Applied Biosystems). Relative mRNA abundance was normalized against Gapdh.
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