Rotor gene q pure detection 2

Manufactured by Qiagen

Sourced in Germany

The Rotor-Gene Q Pure Detection 2.1.0 software is a component of the Rotor-Gene Q real-time PCR system. It provides the core functionality for the operation and analysis of real-time PCR experiments.

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Lab products found in correlation

Rotor gene q, by Qiagen (3 mentions) Genotyping master mix, by Thermo Fisher Scientific (2 mentions) Nuclease free h2o, by Qiagen (2 mentions) Qiaamp dna blood mini kit, by Qiagen (2 mentions) Illustra rnaspin rna isolation kit, by GE Healthcare (1 mentions) Tetro cdna synthesis kit, by Meridian Bioscience (1 mentions) Genotyping assay, by Thermo Fisher Scientific (1 mentions) 2 ml tube, by Eppendorf (1 mentions)

3 protocols using rotor gene q pure detection 2

Gene Expression Analysis in IFNγ, Bradykinin, and LPS-Treated HUVECs

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HUVECs were cultured in 24-well plates to 100% confluency in Comp-MCDB medium, then treated with IFNγ (20 ng/mL), bradykinin (20 μM), or LPS (100 ng/mL) for 2 or 24 h. RNA isolation was carried out using the illustra™ RNASpin RNA Isolation Kit (GE Healthcare, Chicago, CA, USA) according to the manufacturer’s protocol. RNA–cDNA transcription was performed with the Tetro cDNA Synthesis Kit (Bioline, Essex, UK). SensiFAST SYBR Master Mix—No ROX Kit (Bioline) was used for the quantification of cDNA using a Rotor-Gene Q (Qiagen, Hilden, Germany) real-time PCR cycler. Primers (Table 2) were designed using the NCBI Primer-BLAST primer design tool and synthetized using IDT (Coralville, IA, USA). The purity and size of PCR products were checked by sequencing (sequencing was performed by Biomi Ltd., Gödöllő, Hungary) after the first use of each primer pair and by high-resolution melting curve analysis for each measurement.
Quantification was performed using the Rotor-Gene Q Pure Detection 2.1.0 software (Qiagen, Hilden, Germany), and values of interest were normalized with that of β-actin.

Németh Z., Debreczeni M.L., Kajdácsi E., Dobó J., Gál P, & Cervenak L. (2023). Cooperation of Complement MASP-1 with Other Proinflammatory Factors to Enhance the Activation of Endothelial Cells. International Journal of Molecular Sciences, 24(11), 9181.

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DNA Genotyping from Blood and Saliva

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DNA purification from whole blood and saliva samples was performed manually using a QIAamp® DNA Blood Mini Kit (Qiagen Ltd., Manchester, UK), following the manufacturer’s guidelines. DNA samples were then stored at 4°C until subsequent genotyping.
Real-time polymerase chain reaction was performed (Rotor-Gene Q, Qiagen) to establish the genotypes of each SNP for each participant. Each 10 μL reaction volume contained 5 μL Genotyping Master Mix (Applied Biosystems, Foster City, USA), 3.5 μL nuclease-free H₂O (Qiagen), 0.5 μL genotyping assay (Applied Biosystems), plus 1 μL DNA sample. Both negative [1 μL nuclease-free H₂O (Qiagen) replaced the DNA template] and positive controls were included in each RT-PCR run, which used the following protocol: denaturation at 95°C for 10 min, followed by 50 cycles of incubation at 92°C for 15 s, then annealing and extension at 60°C for 1 min. Genotypes were determined using Rotor-Gene Q Pure Detection 2.1.0 software (Qiagen). All samples were analysed in duplicate and there was 100% agreement between genotype calls for samples from the same participant. Genotyping was performed in accordance with published genotyping and quality control recommendations [44 (link)] and all SNPs were genotyped according to the forward DNA strand.

Murtagh C.F., Brownlee T.E., Rienzi E., Roquero S., Moreno S., Huertas G., Lugioratto G., Baumert P., Turner D.C., Lee D., Dickinson P., Lyon K.A., Sheikhsaraf B., Biyik B., O’Boyle A., Morgans R., Massey A., Drust B, & Erskine R.M. (2020). The genetic profile of elite youth soccer players and its association with power and speed depends on maturity status. PLoS ONE, 15(6), e0234458.

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Genotyping of NOS3 Polymorphism from Whole Blood

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A blood sample was drawn into a 10‐ml EDTA vacutainer (BD Vacutainer Systems) from a superficial forearm vein. The whole blood was aliquoted into 2‐ml tubes (Eppendorf AG) and stored at −80°C until subsequent analysis. DNA purification from whole blood samples was performed manually using a QIAamp DNA Blood Mini Kit (Qiagen Ltd.), following the manufacturer's guidelines. DNA samples were then stored at 4°C until subsequent genotyping. Real‐time polymerase chain reaction was performed (Rotor‐Gene Q, Qiagen) to establish the genotypes of each SNP for each participant. Each 10 μl reaction volume contained 5 μl Genotyping Master Mix (Applied Biosystems), 3.5 μl nuclease‐free H₂O (Qiagen), 0.5 μl NOS3 rs2070744 genotyping assay (Applied Biosystems), plus 1 μl DNA sample. Both negative [1 μl nuclease‐free H₂O (Qiagen) replaced the DNA template] and positive controls were included in each RT‐PCR run, which used the following protocol: denaturation at 95°C for 10 min, followed by 50 cycles of incubation at 92°C for 15 s, then annealing and extension at 60°C for 1 min. Genotypes were determined using Rotor‐Gene Q Pure Detection 2.1.0 software (Qiagen). All samples were analyzed in duplicate, and there was 100% agreement between genotype calls for samples from the same participant.

Dawson E.A., Sheikhsaraf B., Boidin M., Erskine R.M, & Thijssen D.H. (2021). Intra‐individual differences in the effect of endurance versus resistance training on vascular function: A cross‐over study. Scandinavian Journal of Medicine & Science in Sports, 31(8), 1683-1692.

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