Quantstudio 3 pcr detection system

Manufactured by Thermo Fisher Scientific

The QuantStudio 3 PCR Detection System is a real-time PCR instrument designed for quantitative gene expression analysis, genotyping, and other molecular biology applications. It features a 96-well format and supports a range of sample volumes and chemistries.

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Lab products found in correlation

Powerup sybr green master mix, by Thermo Fisher Scientific (3 mentions) Maxima first strand cdna synthesis kit for rt pcr, by Thermo Fisher Scientific (3 mentions) Nanodrop nd 1000 spectrophotometer, by Avantor (3 mentions) Mysticq microrna cdna synthesis mix, by Merck Group (2 mentions) Trizol, by Thermo Fisher Scientific (1 mentions)

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QuantStudio 3 (1144 citations) QuantStudio 3 system (282 citations) QuantStudio 3 PCR system (66 citations) QuantStudio 3 detection system (23 citations) QuantStudio 3 real-time PCR detection system (18 citations) QuantStudio PCR Systems (2 citations) QuantStudio™ systems (2 citations)

3 protocols using quantstudio 3 pcr detection system

Quantitative gene expression analysis of iron-related markers

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RNA was isolated using peqGold (Peqlab, Erlangen, Germany) and measured using a Nanodrop ND-1000 spectrophotometer (Peqlab). Reverse transcription was performed with the Maxima First Strand cDNA Synthesis Kit for RT-PCR (Thermo Fisher Scientific, Waltham, USA). RNA expression of IL-1β, IL-8, IL-10, ALOX15, CP, FPN, FTH, FTL, TfR, STEAP3, PHD2, and Glut1 was analyzed using PowerUp SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific) on a QuantStudio 3 PCR Detection System (Applied Biosystems, Thermo Fisher Scientific) and normalized to TBP or 18S. Primers are listed in Table 1.Table 1

List of primers.

Table 1

TARGET	FORWARD (5′-3′)	REVERSE (5′-3′)
ALOX15A	TGGAAGGACGGGTTAATTCTGA	GCGAAACCTCAAAGTCAACTCT
CP	CTTTCCTGCTACCCTGTTTGATGC	CTTGCAAACCGGCTTTCAGA
FTH	TGTGGCGGAGCTGCTGGGTAA	CGAGAGGTGGATACGGCTGCT
FTL	AGCCTTCTTTGTGCGGTCGGGTAA	ACGCCTTCCAGAGCCACATCAT
FPN	TGAGCCTCCCAAACCGCTTCCATA	GGGCAAAAAGACTACAACGACGACTT
Glut1	CACACACCCCTGACACACC	ACTCCCACTGCGACTCTGAC
IL-1Β	AGAAGTACCTGAGCTCGCCA	CAGGTCCTGGAAGGAGCACT
IL-8	CAGGAATTGAATGGGTTTGC	AGCAGACTAGGGTTGCCAGA
IL-10	AAGCCTGACCACGCTTTCTA	TAGCAGTTAGGAAGCCCCAA
PHD2	GAAAGCCATGGTTGCTTGTT	TTGGGTTCAATGTCAGCAAA
STEAP3	CTGCTGTTGCCACCAAAATGC	TTGGCAAGGCTACTATCGCT
TBP	GCATCACTGTTTCTTGGCGT	CGCTGGAACTCGTCTCACTA
TfR	GAGCGTCGGGATATCGGGT	CAGGATGAAGGGAGGACACG
18s	GTAACCCGTTGAACCCCATT	CCATCCAATCGGTAGTAGCG

Schwantes A., Wickert A., Becker S., Baer P.C., Weigert A., Brüne B, & Fuhrmann D.C. (2024). Tumor associated macrophages transfer ceruloplasmin mRNA to fibrosarcoma cells and protect them from ferroptosis. Redox Biology, 71, 103093.

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RNA Expression Analysis of Hypoxia-Related Genes

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RNA was isolated using Trizol (Thermo Fisher Scientific, Waltham, USA) and measured using a Nanodrop ND-1000 spectrophotometer (Peqlab, Erlangen, Germany). Reverse transcription was performed with the Maxima First Strand cDNA Synthesis Kit for RT-PCR (Thermo Fisher Scientific). For Reverse transcription of micro RNA, the MystiCq microRNA cDNA Synthesis Mix (Sigma-Aldrich) was used. RNA expression of in Table 1 listed targets was analyzed using PowerUp SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific) on a QuantStudio 3 PCR Detection System (Applied Biosystems, Thermo Fisher Scientific) and normalized to TBP. Primers are listed in Table 1. SNORD44 primer was purchased from Sigma.Table 1

List of primers

	Forward (5'–3')	Reverse (5'–3')
TBP	GCATCACTGTTTCTTGGCGT	CGCTGGAACTCGTCTCACTA
PPTC7	CGAGGGAGTCGTCTTGAGC	GCCGTCAGGATAATGTCTCCTA
HIF-1α	GCTGGCCCCAGCCGCTGGAG	GAGTGCAGGGTCAGCACTAC
HIF-2α	AAGCCTTGGAGGGTTTCATT	TGCTGGATTGGTTCACACAT
ARNT	ACTACTGCCAACCCCGAAAT	CTCTGGACAATGGCTCCTCC
GLUT1	TCACTGTGCTCCTGGTTCTG	CCTGTGCTCCTGAGAGATCC
HK2	GTGAATCGGAGAGGTCCCAC	GCTAACTTCGGCCACAGGAT
PFKL	TCGACTGCAGGACCAATGTC	AGCTTCTCCGACAACCACAG
LDHA	ACGTCAGCAAGAGGGAGAAA	CGCTTCCAATAACACGGTTT
PFKFB1	GAAACCCAGTACACCCCCTG	TTCTGCAACCTGGTTTGGGT
PFKFB2	AGGCAGGGAGGGATCTTAGG	CGATCAGAGTCGGGGAGTTG
PFKFB3	CAGCTGCCTGGACAAAACAT	GAGGGCAGGACACAAGCTAA
PFKFB4	GGGTGCCTCTTGGCCTTAAA	GCCCACACGGCATACTTTTC
miR-193a-3p	AACTGGCCTACAAAGTCCCAGT	MystiCq Universal Primer

Fuhrmann D.C, & Brüne B. (2022). miR-193a-3p increases glycolysis under hypoxia by facilitating Akt phosphorylation and PFKFB3 activation in human macrophages. Cellular and Molecular Life Sciences, 79(2), 89.

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Comprehensive Analysis of Iron Metabolism Genes

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RNA was isolated using peqGold (Peqlab, Erlangen, Germany) and measured using a Nanodrop ND-1000 spectrophotometer (Peqlab). Reverse transcription was performed with the Maxima First Strand cDNA Synthesis Kit for RT-PCR (Thermo Fisher Scientific, Waltham, USA). For Reverse transcription of micro RNA the MystiCq microRNA cDNA Synthesis Mix (Sigma-Aldrich) was used. RNA expression of DMT1, TfR, CP, FPN, FTH, FTMT, FTL, HERC2, PCBP1, NCOA4, and miR-6862-5p was analyzed using PowerUp SYBR Green Master Mix (Applied Biosystems, Thermo Fisher Scientific) on a QuantStudio 3 PCR Detection System (Applied Biosystems, Thermo Fisher Scientific) and normalized to TBP. Primers are listed in Table 1. SNORD44 primers were purchased from Sigma.Table 1

List of primers.

Table 1

	Forward (5′-3′)	Reverse (5′-3′)
TBP	GCATCACTGTTTCTTGGCGT	CGCTGGAACTCGTCTCACTA
DMT1	AGCCACTCAGGTATCCACCAT	CCAGGGGACTGTGAAAGAGAG
TfR	GAGCGTCGGGATATCGGGT	CAGGATGAAGGGAGGACACG
CP	CTTTCCTGCTACCCTGTTTGATGC	CTTGCAAACCGGCTTTCAGA
FPN	TGAGCCTCCCAAACCGCTTCCATA	GGGCAAAAAGACTACAACGACGACTT
FTH	TGTGGCGGAGCTGCTGGGTAA	CGAGAGGTGGATACGGCTGCT
FTL	AGCCTTCTTTGTGCGGTCGGGTAA	ACGCCTTCCAGAGCCACATCAT
FTMT	CATGCCATGGAGTGTGCTCT	AATCGCACAAATGGGGGTCA
HERC2	TGCTTCCTACTTAGGCGTGC	TGGCAAGAGAAGGGCGATTT
PCBP1	TCGGCTTCTTATGCACGGAA	GGCCGGTCAGAGTGATGATT
NCOA4	TCAAGATGTAACCGTTGGGAA	AGCAGAAAGGCTGCTCAACT
miR-6862-5p	CGGGCATGCTGGGAGAGACTTT	MystiCq Universal Primer

Fuhrmann D.C., Mondorf A., Beifuß J., Jung M, & Brüne B. (2020). Hypoxia inhibits ferritinophagy, increases mitochondrial ferritin, and protects from ferroptosis. Redox Biology, 36, 101670.

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