RNA was extracted using Trizol (Invitrogen, Carlsbad, MA, USA), with 1 μg reverse transcribed using qScript Supermix Reagent (Quanta Biosciences, Gaithersburg, MD, USA). Primers for α2M were forward 5′-CCAGGACACGAAGAAGG-3′ and reverse 5′-CACTTCACGATGAGCAT-3′. Quantitative PCR was performed using the Power SYBR Green (Applied Biosystems, Waltham, MA, USA) PCR Master Mix on the Vii 7 Real-Time PCR System (Applied Biosystems, Waltham, MA, USA). Changes in mRNA expression were determined relative to 18S using the ΔΔCt method.
Qscript supermix reagent
Manufactured by Quanta Biosciences
Sourced in United States
The QScript Supermix Reagent is a ready-to-use, high-performance solution for reverse transcription and real-time PCR. The reagent contains all the necessary components, including reverse transcriptase, RNase inhibitor, and optimized buffer, to facilitate efficient and reliable gene expression analysis.
Automatically generated - may contain errors
Lab products found in correlation
4 protocols using qscript supermix reagent
1
Quantitative RT-PCR Analysis of α2M
2
Quantifying Kidney Stress Responses
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RNA was extracted from flash-frozen kidney sections using the RNeasy Plus Mini Kit (Qiagen). RNA was reverse transcribed using qScript Supermix Reagent (Quanta Biosciences) and quantitative real time PCR was performed using the Power SYBR Green PCR Master Mix on the Applied Biosystems Vii 7 Real-Time PCR System. Primer sequences used were as follows: CHOP forward 5′-GGAAACAGAGTGGTCATTCCC-3′ and reverse 5′-CTGCTTGAGCCGTTCATTCTC-3′ and IRE1α forward 5′-CCATCGAGCTGTGTGCAG-3′ and reverse 5′-TGTTGAGGGAGTGGAGGTG-3′. mRNA expression levels were determined relative to 18S of the same sample using the ΔΔCT method.
3
Quantifying TGFβ1 mRNA Expression
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Trizol (Invitrogen) was used to extract RNA and 1 µg was reverse transcribed using qScript Supermix Reagent (Quanta Biosciences). Using Power SYBR Green PCR Master Mix on the Applied Biosystems Vii 7 Real-Time PCR System, quantitative PCR was performed to determine changes in mRNA expression relative to 18 S using the ΔΔCt method with the following primers: TGFβ1 forward 5′-AAACGGAAGCGCATCGAA-3′ and reverse 5′-GGGACTGGCGAGCCTTAGTT-3′ and 18S forward 5′-GCCGCTAGAGGTGAAATTCTTG-3′ and reverse 5′-CATTCTTGGCAAATGCTTTCG-3′.
4
Quantitative Analysis of TSP1 mRNA Expression
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RNA was extracted from MC using Trizol (Invitrogen), and 0.5 µg of RNA was reverse transcribed using qScript Supermix Reagent (Quanta Biosciences). Expression of TSP1 mRNA relative to 18S was determined using the ΔΔCt method. Quantitative PCR was performed using Power SYBR Green PCR Master Mix on the Applied Biosystems Vii 7 Real-Time PCR System. The following primers were used: TSP1 forward 5′-TGGCCAGCGTTGCCA -3′ and reverse 5′- TCTGCAGCACCCCCTGAA-3′ and 18S forward 5′- GCCGCTAGAGGTGAAATTCTTG-3′ and reverse 5′- CATTCTTGGCAAATGCTTTCG-3’.
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