Icycler iq real time pcr instrument

Manufactured by Bio-Rad

Sourced in Canada

The iCycler iQ real-time PCR Instrument is a thermal cycler designed for real-time polymerase chain reaction (PCR) analysis. It is capable of detecting and quantifying nucleic acid sequences in real-time during the amplification process.

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Lab products found in correlation

Oligo dt, by Thermo Fisher Scientific (1 mentions) Sybr green mixture, by Bio-Rad (1 mentions) Qiashredder, by Qiagen (1 mentions) Quantitect sybergreen, by Qiagen (1 mentions) Microrneasy kit, by Qiagen (1 mentions) Super script 3 reverse transcriptase, by Merck Group (1 mentions) Iq sybr green supermix, by Bio-Rad (1 mentions)

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ICycler (1160 citations) ICycler iQ (245 citations) ICycler instrument (23 citations) ICycler iQ instrument (10 citations) ICycler real-time PCR instrument (9 citations) ICycler PCR instrument (8 citations)

4 protocols using icycler iq real time pcr instrument

Real-Time PCR Gene Expression Analysis

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Total RNA isolation and reverse transcription with oligo(dT) (18418–012, Invitrogen) were performed as described previously [14 (link)]. The amounts of individual genes were measured with gene-specific primers by real-time PCR analysis with a iCycler iQ real-time PCR Instrument (Bio-Rad) and SYBR Green mixture (Bio-Rad). The relative expression or amount of specific genes was quantitated with the 2^-ΔΔCt calculation [15 (link)], according to the manufacturer’s software (Bio-Rad), where the reference gene was ubiquitin. Primers used in real-time RT-PCR are in Additional file 1.

Ding Y., Virlouvet L., Liu N., Riethoven J.J., Fromm M, & Avramova Z. (2014). Dehydration stress memory genes of Zea mays; comparison with Arabidopsis thaliana. BMC Plant Biology, 14, 141.

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Real-Time PCR Analysis of Hexose Transporters

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Real-time PCR on the expression of HXT1-17 and GAL2 was performed with the primers indicated in Additional file 1: Table S3, using the SensiMix SYBR & Fluorescein Kit (Quantace Ltd.) and the iCycler iQ Real-Time PCR instrument (Bio-Rad). Actin was used as a reference gene to normalize fold changes. The SYBR Green Master Mix (12.5 μl) was used for 25 μl reactions containing 4 μl of the extracted total RNA (10 ng/μl), 1 μl of the indicated primers (10 nM), and 6.5 μl of sterile water. The PCR conditions for HXT1-17 and GAL2 were 10 min at 95°C followed by 40 cycles of amplification (15 s at 95°C, 30 s at 58°C, 30 s at 72°C).

Nijland J.G., Shin H.Y., de Jong R.M., de Waal P.P., Klaassen P, & Driessen A.J. (2014). Engineering of an endogenous hexose transporter into a specific D-xylose transporter facilitates glucose-xylose co-consumption in Saccharomyces cerevisiae. Biotechnology for Biofuels, 7, 168.

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Quantitative Real-Time PCR Analysis of Lymphatic Vessels

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Following dissection and incubation procedures similar to those described above, lymphatic vessels were placed in RNA-later (Thermo Fisher Sientific) for 24 h, homogenized using a QIAshredder (Qiagen, Mississauga, ON, Canada) and RNA extracted using the Qiagen Micro RNeasy Kit. The extracted RNA was used to synthesize cDNA with SuperscriptⅡReverse Transcriptase enzyme and oligo (dT) primers (both from Thermo Fisher Sientific) and the target genes (β-actin, eNOS, iNOS, COX-1, COX-2, TNF-α and TNFR1) were amplified by using QuantiTect Syber Green (Qiagen) in an iCycler iQ Real-Time PCR Instrument (Bio-Rad, Mississauga, ON, Canada). The sequences of all primers were determined according to the NCBI published rat sequences and forward and reverse primers were designed using Primer3 Software [39 (link),66 (link)] (see Table 1). PCR results were quantified using the ΔΔCT method [81 (link)], and mRNA expression of the target genes was normalized to that of β-actin.

Chen Y., Rehal S., Roizes S., Zhu H.L., Cole W.C, & von der Weid P.Y. (2017). The pro-inflammatory cytokine TNF-α inhibits lymphatic pumping via activation of the NF-κB - iNOS signaling pathway. Microcirculation (New York, N.Y. : 1994), 24(3), 10.1111/micc.12364.

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Quantitative Real-Time PCR Analysis

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cDNA was synthesized using Superscript III Reverse Transcriptase (Sigma) according to the manufacturer’s instructions. qPCR was performed in 96-well plates using iQ™ SYBR Green Supermix (Bio-Rad) and an I-cycler iQ Real-Time PCR instrument (Bio-Rad). Measurements were performed in triplicate with a variability <0.5 Ct. mRNA expression was analysed by normalizing to that of the housekeeping gene GADPH. Primers were designed with PERL primer software using NCBI EntrezGene reference sequences as templates and synthesized by Sigma. The primers used are listed in Supplementary Table S4.

Arcidiacono P., Ragonese F., Stabile A., Pistilli A., Kuligina E., Rende M., Bottoni U., Calvieri S., Crisanti A, & Spaccapelo R. (2017). Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells. European Journal of Nutrition, 57(7), 2547-2569.

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