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Icycler rt pcr detection system

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The iCycler RT-PCR detection system is a real-time PCR instrument designed for quantitative gene expression analysis. It provides thermal cycling, fluorescence detection, and data analysis capabilities to enable researchers to perform real-time reverse transcription PCR (RT-PCR) experiments.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (15 mentions) Sybr premix ex taq 2, by Takara Bio (10 mentions) Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (9 mentions) Rneasy plus kit, by Qiagen (8 mentions) Sybr green master mix, by Bio-Rad (7 mentions) Cdna synthesis kit, by Bio-Rad (6 mentions) Tri reagent, by Thermo Fisher Scientific (4 mentions) Trizol, by Thermo Fisher Scientific (3 mentions) Iq sybr green supermix, by Bio-Rad (3 mentions) Primescript rt reagent kit, by Takara Bio (2 mentions) Rneasy micro kit, by Qiagen (2 mentions) Dnase 1, by Thermo Fisher Scientific (2 mentions) Chamq universal sybr qpcr master mix, by Vazyme (2 mentions) Zymolyase 20t, by MP Biomedicals (2 mentions) Iscript cdna synthesis kit, by Bio-Rad (2 mentions) Rnaiso plus, by Takara Bio (1 mentions) Gdna eraser, by Takara Bio (1 mentions) M mlv, by Promega (1 mentions) Primescript rt master mix, by Takara Bio (1 mentions) Chamq sybr qpcr master mix, by Vazyme (1 mentions)

Close spelling variants of this product

ICycler (1160 citations) ICycler system (143 citations) RT-PCR system (39 citations) RT-PCR detection system (9 citations) ICycler RT-PCR system (7 citations) ICycler iQ RT-PCR detection system (7 citations) ICycler iQ5 multicolor RT-PCR detection system (2 citations) ICycler iQ5 RT-PCR detection system (2 citations)

30 protocols using icycler rt pcr detection system

1

Quantification of HUWE1 mRNA Levels

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Total RNA was extracted from whole blood using RNAiso Plus (D9108A; TaKaRa, Japan). Total RNA (500 ng) was used for reverse transcription with a PrimeScriptRT reagent kit and gDNA Eraser (DRR047A; TaKaRa). Quantitative PCR (qPCR) was performed using SYBR Premix Ex Taq II (RR820A, TaKaRa) on an iCycler RT-PCR detection system (Bio-Rad, CA) to analyze the relative mRNA expression levels of HUWE1. The ΔΔCT method was used for data analysis. Each assay was performed in triplicate for each sample.
Absolute quantitation of HUWE1 copy numbers was performed with two independent tests, using GAPDH and α-tubulin as internal controls. The primers for RT-PCR are listed in Supplementary Materials Table S1.
Zebra sh embryo RNA isolation and real-time PCR (RT-PCR) RNA was isolated from a pool of embryos at different stages using TRIzol reagent (Gibco BRL, USA).
Total RNA (1 µg) was reverse transcribed. The relative mRNA expression levels of huwe1 were ampli ed using SYBR Premix Ex Taq II (RR820A, TaKaRa) on an iCycler RT-PCR detection system (Bio-Rad, CA). The gapdh gene was used as an internal control. The primers for RT-PCR are listed in Supplementary Materials Table S1. After 40 cycles, ampli ed cDNAs were analyzed on agarose gels.
Wu H., Liang Y., Li J., Zhang X., Xu B., Wang X., Cheng J., Dinnyés A., Xie J., & Xu W. (2021). A significant effect of the HUWE1 copy number on autistic spectrum disorder and the development of zebrafish.
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2

RNA Isolation and qRT-PCR Analysis

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Fixed cochleae, whole organoids from a single well, and sorted cells were lysed into RLT buffer and total RNA was isolated using the RNeasy Micro Kit (QIAGEN) according to the manufacturer's instructions. RNA was converted to cDNA with the qScript XLT cDNA kit (Quanta BioSciences). qRT-PCR was performed with diluted cDNA (1:4) in two wells for each primer and TaqMan probes and SYBR green master mix (Bio-Rad) on the Bio-Rad iCycler RT-PCR detection system.
Abdul-Aziz D., Hathiramani N., Phung L., Sykopetrites V, & Edge A.S. (2021). HIC1 Represses Atoh1 Transcription and Hair Cell Differentiation in the Cochlea. Stem Cell Reports, 16(4), 797-809.
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3

Real-Time qPCR Protocol for Gene Expression

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Total RNA was extracted from mouse tissues or cells using TRIzol reagent (Invitrogen) and converted to cDNA using a RevertAid First-Strand cDNA Synthesis Kit (ThermoFisher). Real-time PCR was performed using SYBR Premix Ex Taq II (TaKaRa, Dalian, China) on an iCycler RT-PCR Detection System (Bio-Rad Laboratories, Hercules, USA). The ΔΔCT method was used for data analysis. Each assay was performed in triplicate for each sample. The
actin gene was used as an internal control. The primers for real-time PCR are listed in
Supplementary Table S1.
Wu S., Li X., Shang L., Wu L., Li T., Li P., Ji Z., Hou J., Yin M, & Xu W. (2022). The novel BRDT inhibitor NHWD870 shows potential as a male contraceptive in mice: A novel inhibition of BRDT for male contraception. Acta Biochimica et Biophysica Sinica, 54(12), 1789-1800.
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4

RT-qPCR Protocol for Exosomal lncRNA Analysis

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Tissues, cells or exosomes were lysed in Trizol Reagent (Invitrogen) and total RNA was isolated according to the manufacturer’s instructions with following modification for exosomal RNA purification: the aqueous phase containing total RNA was purified using the RNeasy plus kit (Qiagen). RNA was converted to cDNA with the cDNA synthesis kit (M-MLV, Promega). RT-qPCR was performed with diluted cDNA (1:10) in three wells for each primer and SYBR green master mix (Bio-Rad) on Bio-Rad iCycler RT-PCR detection system. All RT-qPCR experiments were repeated at least three independent times. Relative expression was calculated with 2−ΔΔCt., using β-actin as a reference gene and compared with the indicated control sample(s). Housekeeping gene β-actin was also used as a reference gene for exosomal lncRNAs according to previous literatures43 (link)44 (link)45 (link), for the following reasons. On one hand, lncRNAs, specifically Hotair are RNAs with polyA end, and thus only the RNAs with polyA are suitable for reference. On the other hand, it seems that the genes abundantly expressed in cells have the trend to be present at levels comparable to its cellular levels. Primers used are listed on Supplementary Table 1.
Lu X., Bai D., Liu X., Zhou C, & Yang G. (2017). Sedentary lifestyle related exosomal release of Hotair from gluteal-femoral fat promotes intestinal cell proliferation. Scientific Reports, 7, 45648.
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5

Quantitative Real-Time PCR Analysis of Mouse Tissue Transcripts

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Total RNA was extracted from mouse tissues using TRIzol reagent (Invitrogen) and was reverse transcribed into cDNA by PrimeScript RT Master Mix (Takara). ChamQ SYBR qPCR Master Mix (Vazyme) was utilized to carry out real-time PCR on an iCycler RT‒PCR Detection System (Bio-Rad Laboratories). Data analysis was performed using the ΔΔCT method. Each sample was repeated three times for each assay. The Gapdh gene was used as an internal control. Table S5 contains a list of the real-time PCR primers.
Wu Y., Li J., Li C., Lu S., Wei X., Li Y., Xia W., Qian C., Wang Z., Liu M., Gu Y., Huang B., Tan Y, & Hu Z. (2023). Fat mass and obesity-associated factor (FTO)-mediated N6-methyladenosine regulates spermatogenesis in an age-dependent manner. The Journal of Biological Chemistry, 299(6), 104783.
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6

Quantitative Real-Time PCR Protocol for Mouse

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Total RNA of mice tissues was extracted using TRIzol reagent (Invitrogen) and was converted to cDNA using a RevertAid First-Strand cDNA Synthesis Kit (ThermoFisher). Real-time PCR was performed using SYBR Premix Ex Taq II (TaKaRa) on an iCycler RT-PCR Detection System (Bio-Rad Laboratories). The ΔΔCT method was used for data analysis. Each assay was performed in triplicate for each sample. The Gapdh gene was used as an internal control. The primers for real-time PCR are listed in Supplementary Table S5.
Lu S., Gu Y., Wu Y., Yang S., Li C., Meng L., Yuan W., Jiang T., Zhang X., Li Y., Wang C., Liu M., Ye L., Guo X., Shen H., Yang X., Tan Y, & Hu Z. (2021). Bi-allelic variants in human WDR63 cause male infertility via abnormal inner dynein arms assembly. Cell Discovery, 7, 110.
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7

RNA Extraction, Conversion, and qRT-PCR Analysis

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Total RNA was isolated from mESCs using RNeasy plus kit (Qiagen) or Trizol (Invitrogen), and treated with DNaseI (Life Technologies) to remove the DNA contamination. RNA was converted to cDNA with a cDNA synthesis kit (Bio-Rad). qRT-PCR was performed with SYBR green master mix (Bio-Rad) on Bio-Rad iCycler RT-PCR detection system according to manufacture’s instructions. Small RNA RT-PCR was performed using Taqman miRNA assays (Life Technologies) as described before (Das et al., 2008 (link)) .
Das P.P., Hendrix D.A., Apostolou E., Buchner A.H., Canver M.C., Beyaz S., Ljuboja D., Kuintzle R., Kim W., Karnik R., Shao Z., Xie H., Xu J., De Los Angeles A., Zhang Y., Choe J., Jun D.L., Shen X., Gregory R.I., Daley G.Q., Meissner A., Kellis M., Hochedlinger K., Kim J, & Orkin S.H. (2015). PRC2 is required to maintain expression of the maternal Gtl2-Rian-Mirg locus by preventing de novo DNA methylation in mouse embryonic stem cells. Cell reports, 12(9), 1456-1470.
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8

Quantitative RT-PCR Analysis of Wnt Signaling

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Total RNA was isolated from tumours or cells using the RNeasy plus kit (Qiagen) according to the manufacturer’s instructions. cDNA was synthesized from 1 μg of RNA using the SuperScript VILO cDNA synthesis kit (Thermo). qPCR was performed in triplicates with 2 μl of diluted cDNA (1:10) using PerfeCTa SYBR Green FastMix (Thermo) on a Bio-Rad iCycler RT–PCR detection system. Expression was normalized to Actb or Gapdh. All oligonucleotides used in this study are listed in Supplementary Data Table 4. All qPCR experiments were reproduced using at least three biological replicates.
Alternatively, a Mouse WNT Signaling Pathway RT2 Profiler PCR Array (Qiagen) was used according to manufacturer’s instructions. Raw expression values were thresholded to drop non-detected and lowly expressed genes (maximum Ct value set to 33; 0 values set to 33). Array position to gene-name mapping details were retrieved from the manufacturer’s website (www.pcrdataanalysis.sabiosciences.com). Expression values for all genes per array were normalized to the expression of the housekeeping gene Gusb. Three replicates of stroma samples and three replicates of tumour samples were compared to calculate log2 fold-change and differential expression significance values (2-sided t-test).
Tammela T., Sanchez-Rivera F.J., Cetinbas N.M., Wu K., Joshi N.S., Helenius K., Park Y., Azimi R., Kerper N.R., Wesselhoeft R.A., Gu X., Schmidt L., Cornwall-Brady M., Yilmaz Ö.H., Xue W., Katajisto P., Bhutkar A, & Jacks T. (2017). A Wnt-producing niche drives proliferative potential and progression in lung adenocarcinoma. Nature, 545(7654), 355-359.
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9

Quantitative RT-PCR Analysis of Gene Expression

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Total RNA was isolated from cell lines and tissues using a Super-Purity RNA Exaction Kit (BioTeke). First-strand cDNA was reverse transcribed using a RevertAid First-Strand cDNA Synthesis Kit (ThermoFisher, Waltham, MA). The primers that were used for RT-PCR and qRT-PCR are listed in supplementary Table S2. The GAPDH gene was used as an internal control.
Quantitative RT-PCR was performed using the SYBR Premix Ex Taq II (TaKaRa Bio). Reactions were run using a Bio-Rad iCycler RT-PCR Detection System. The ΔΔCT method was used for data analysis. Each assay was performed in triplicate using each sample.
Lu Y., Liu Y., Liao S., Tu W., Shen Y., Yan Y., Tao D., Lu Y., Ma Y., Yang Y, & Zhang S. (2016). Epigenetic modifications promote the expression of the orphan nuclear receptor NR0B1 in human lung adenocarcinoma cells. Oncotarget, 7(28), 43162-43176.
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10

Yeast Starvation RNA Extraction and qRT-PCR

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Yeast cells were grown in YPD to log phase. Then, Yeast cells were collected and washed twice with distilled water and cultured in SD‐N media for the indicated time. Cells were collected and frozen in liquid nitrogen. Cell walls were lysed with the ZYMOLYASE‐20T (MP Biomedicals, 320921) and then FastPure cell/tissue total RNA isolation mini kit (Vazyme, RC101‐01) was used to extract total RNA. Extracted RNA was converted to cDNA using HiScript III RT SuperMix for qPCR (Vazyme, R323‐01) and qRT‐PCR reactions were performed using ChamQ Universal SYBR qPCR Master Mix (Vazyme, C711‐02) on an iCycler RT‐PCR Detection System (Bio‐Rad Laboratories). Each assay was performed in triplication for each sample. The reaction mix is in a 15‐μl final volume consisting of 7.5 μl of SYBR Green Master mix, 0.5 μl of each primer with 333.3 nM final concentration, 1.5 μl of H2O, and 5 μl of the cDNA preparation. The thermocycling program is 95°C for 10 min, 40 cycles of 95°C for 15 s, and 60°C for 1 min. After that, melting curves were analyzed to verify PCR specificity. The transcript abundance was analyzed by a comparative threshold‐cycle method. The relative abundance of the reference mRNAs of TFC1 was analyzed and taken for normalization of differences in total RNA amount.
Huang T., Jiang G., Zhang Y., Lei Y., Liu S., Li H, & Lu K. (2022). The RNA polymerase II subunit Rpb9 activates ATG1 transcription and autophagy. EMBO Reports, 23(11), e54993.
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