Pikoreal

Manufactured by Thermo Fisher Scientific

Sourced in United States, Japan

The PikoReal is a compact, real-time PCR (polymerase chain reaction) instrument designed for high-performance nucleic acid detection and quantification. It features a 96-well format and supports a wide range of sample volumes and reaction chemistries. The PikoReal enables precise, sensitive, and reproducible real-time PCR analysis across a variety of applications.

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Lab products found in correlation

Trizol reagent, by Thermo Fisher Scientific (3 mentions) Primescript rt reagent kit, by Takara Bio (2 mentions) Maxima sybr green rox qpcr master mix, by Thermo Fisher Scientific (2 mentions) Primescript rt master mix, by Takara Bio (2 mentions) One step sybr reverse transcription pcr rt pcr kit, by Takara Bio (2 mentions) Thunderbird sybr qpcr mix, by Toyobo (2 mentions) Fluorescence quantitative pcr kit, by Takara Bio (1 mentions) Cellulose nitrate membrane, by Merck Group (1 mentions) Sybr premix ex taq, by Takara Bio (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Taqman probe, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Bio-Rad (1 mentions) Trizol reagent, by Takara Bio (1 mentions) One step sybr rt pcr kit, by Takara Bio (1 mentions) Rna solv reagent, by Omega Bio-Tek (1 mentions) Dnase, by Thermo Fisher Scientific (1 mentions) High capacity archive kit, by Thermo Fisher Scientific (1 mentions) Tb green premix ex taq 2, by Takara Bio (1 mentions) Trizol, by CWBIO (1 mentions) One step sybr primescript rt pcr kit 2, by Takara Bio (1 mentions)

Close spelling variants of this product

PikoReal 96 Real-Time PCR System (125 citations) PikoReal Software 2.2 (15 citations) PikoReal software (14 citations) PikoReal Real-Time PCR (13 citations) PikoReal RT-PCR system (11 citations) PikoReal Real-Time PCR machine (10 citations) PikoReal 96 system (6 citations) PikoReal Thermocycler (4 citations) PikoReal 96 machine (4 citations) PIKOREAL 96 fluorescence quantitative PCR instrument (3 citations)

21 protocols using pikoreal

Quantifying Bcl-2 and Bax mRNA Expression

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A PCR kit (PikoReal; Thermo Fisher Scientific, Inc., Waltham, MA, USA) was used to detect the expression of Bcl-2 mRNA and Bax mRNA in left kidney tissues. TRIzol was used to extract total RNA [(Invitrogen, Inc., Carlsbad, CA, USA); fluorescence quantitative PCR kit (Takara Bio, Inc., Otsu, Japan); cellulose nitrate membrane (Millipore Corp., Billerica, MA, USA)] according to the manufacturer's instructions. After RNA extraction, UV spectrophotometry was used to measure the OD values at 260 and 280 nm. The RNA sample was stored at −80°C for later use.
Reverse transcription of RNA to cDNA was performed according to the instructions of the reverse transcription kit. RNAase-free water was used to dilute the product 10 times. For fluorescence quantitative PCR amplification, the Livak method (2^−ΔΔCt) was used for relative quantitation (β-actin as control). Bcl-2 gene was amplified with: Forward, 5′-GTGGTGGAGGAACTCTTCAGGGATG-3′ and reverse, 5′-GGTCTTCAGAGACAGCCAGGAGAAATC-3′ (226 bp); Bax gene was amplified with: Forward, 5′-GGGTTTCATCCAGGATCGAGCAG-3′ and reverse, 5′-GAGTCCGTGTCCACGTCAGCAAT-3′ (288 bp); and β-actin gene was amplified with: Forward, 5′-ATGTGGCCGAGGACTTTGATT-3′ and reverse, 5′-AGTGGGGTGGCTTTTAGGATG-3′ (107 bp).

Xia Q., Liu C, & Zheng X. (2017). N-acetylcysteine ameliorates contrast-induced kidney injury in rats with unilateral hydronephrosis. Molecular Medicine Reports, 17(2), 2203-2210.

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Transcriptional Analysis of Murine Tibial RNA

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Total RNA was extracted from the left tibia tissue of mice to perform RNA reverse transcription. The cycle threshold (Ct) of the samples detected during PCR was analysed by Thermo Scientific PikoReal (Thermo). The relative mRNA expression level was calculated by the 2^-△△CT method.

Xu D., Gao H.J., Lu C.Y., Tian H.M, & Yu X.J. (2022). Vitamin D inhibits bone loss in mice with thyrotoxicosis by activating the OPG/RANKL and Wnt/β-catenin signaling pathways. Frontiers in Endocrinology, 13, 1066089.

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Quantification of Aortic ET-1 mRNA Expression in Rats

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SYBR-Green RT-qPCR was performed in order to detect the mRNA levels of ET-1. Total RNA was extracted from the aortic tissue of rats using TRIzol reagent (Invitrogen, Grand Island, NY, USA). RNA was reverse transcribed to cDNA using the PrimeScript RT reagent kit (Takara Bio, Otsu, Japan). qPCR was performed with PikoReal (Thermo Fisher Scientific, Waltham, MA, USA) using SYBR Premix Ex Taq (Takara Bio). The relative quantification values for the expression of these genes were calculated using the ΔΔCT method and corrected using a housekeeping gene. The following primer sequences for the analysis of ET-1 and GAPDH mRNA were used: ET-1 forward, 5′-CAACCAGACACCGTCCTCTT-3′ and reverse, 5′-CTTGGAAAGCCACAAACAGC-3′; and GAPDH forward, 5′-TCATTGACCTCAACTACA-3′ and reverse, 5′-CAAAGTTGTCATGGATGACC-3′.

TANG S.T., SU H., ZHANG Q., TANG H.Q., WANG C.J., ZHOU Q., WEI W., ZHU H.Q, & WANG Y. (2016). Sitagliptin inhibits endothelin-1 expression in the aortic endothelium of rats with streptozotocin-induced diabetes by suppressing the nuclear factor-κB/IκBα system through the activation of AMP-activated protein kinase. International Journal of Molecular Medicine, 37(6), 1558-1566.

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Quantification of Metabolic Gene Expression

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Total RNA was extracted from J774A.1 cells for each group in triplicate using Qiagen's RNeasy Mini Kit and was reverse transcribed using iScript cDNA Synthesis Kit (BioRad Laboratories, Hercules, CA, USA). Polymerase chain reaction was performed using Piko-Real (Thermofisher, Fremont CA, USA) for RNA expression levels of genes of interest, namely, lactate dehydrogenase A and B isoforms (LDHA/B) and monocarboxylate transporters 1 and 4 isoforms (MCT1/4), using Taqman probes (Applied Biosystems, Foster City, CA, USA) as described previously 11 (link). β-actin was used as the housekeeping gene, and the relative difference from the control cells was calculated for each primer/probe combination.

Sriram R., Nguyen J., Santos J.D., Nguyen L., Sun J., Vigneron S., Van Criekinge M., Kurhanewicz J, & MacKenzie J.D. (2018). Molecular detection of inflammation in cell models using hyperpolarized 13C-pyruvate. Theranostics, 8(12), 3400-3407.

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Compound 9 Modulates LPS-Induced Responses

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RAW264.7 cells were placed in a 6-well plate at a density of 1 × 10⁶ cells/well and incubated for 12 h. Cultured cells were pretreated with compound 9 (1, 2, 4 μM/L) for 1 h and incubated with LPS (1 μg/mL) for 12 h. RNA was extracted with Trizol Reagent (Takara Bio Inc., Otsu, Japan) according to the manufacturer’s instructions, and cDNA was reverse transcribed from total RNA using a Superscript III system (Takara Bio Inc., Otsu, Japan). PCR amplification was carried out using PikoReal™ (Thermo Fisher Scientific, MA, US) and specific primers. The primer sequences (Wcgene Biotech, Shanghai, China) are shown in Table 3. The optimal conditions for PCR amplification of the cDNA were established by following the manufacturer’s instructions. Relative gene expression was calculated using the comparative Ct method (2^−ΔΔCt) with glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as an internal control. All experiments were performed in triplicate (n = 3).

Liu H., Yan C., Li C., You T, & She Z. (2020). Naphthoquinone Derivatives with Anti-Inflammatory Activity from Mangrove-Derived Endophytic Fungus Talaromyces sp. SK-S009. Molecules, 25(3), 576.

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Quantitative RT-PCR for AMIGO2 and GAPDH

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The human AMIGO2 RT-PCR forward and reverse primers were 5′-GATACTGCAGCAGGGCAGAA-3′ and 5′-GACGCCACAAAAGGTGTGTC-3′, respectively. The forward and reverse murine AMIGO2 primers were 5′-GGCACTTTAGCTCCGTGATG-3′ and 5′-GTCTCGTTTAACAGCCGCTG-3′, respectively, as previously described (Kuja-Panula et al., 2003 (link)). For the mouse GAPDH control, the forward and reverse primers were 5′-CAACGACCCCTTCATTGACC-3′ and 5′-AGTGATGGCATGGACTGTGG-3′, respectively. Quantitative real-time PCR was performed in a real-time PCR system (PikoReal; Thermo Fisher Scientific).

Park H., Lee S., Shrestha P., Kim J., Park J.A., Ko Y., Ban Y.H., Park D.Y., Ha S.J., Koh G.Y., Hong V.S., Mochizuki N., Kim Y.M., Lee W, & Kwon Y.G. (2015). AMIGO2, a novel membrane anchor of PDK1, controls cell survival and angiogenesis via Akt activation. The Journal of Cell Biology, 211(3), 619-637.

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Quantifying mRNA Expression in Liver Samples

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The relative mRNA expression levels were measured using real-time RT-PCR. The total liver RNA was extracted using RNA-Solv Reagent (Omega Bio-Tek Inc., Norcross, GA, USA). The concentration of each sample was processed to 50 ng/μl with DEPC water using a spectrophotometer (Thermo Fisher Scientific K.K., Tokyo, Japan). The RNA was reverse-transcribed and amplified using the One-Step SYBR RT-PCR kit (Takara Bio Inc., Shiga, Japan) with primers for target genes on a real-time RT-PCR system (PikoReal, Thermo Fisher Scientific, Waltham, MA, USA). The relative mRNA expression levels of the target genes were corrected using Gapdh and were analyzed using the ΔΔCt method.

Ichikawa N., Ng L.S., Makino S., Goh L.L., Lim Y.J., F., Sasaki H., Shibata S, & Lee C.L. (2022). Solid-State Fermented Okara with Aspergillus spp. Improves Lipid Metabolism and High-Fat Diet Induced Obesity. Metabolites, 12(3), 198.

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Quantitative PCR Analysis of Immune Genes

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Collection of blood samples, PBMC isolation, and RNA extraction were carried out according to previously described protocols (Chase et al., 2015 (link)). RNA extracts were treated with DNAse (Ambion) to remove any possible genomic DNA contaminants, and reverse transcribed using the Applied Biosystems High Capacity Archive Kit. Maxima SYBR Green/ROX qPCR Master Mix (#K0222) was used for detection of PCR product and mixtures were run on a Thermo Scientific™ PikoReal. Relative quantification values were calculated using the delta delta ct method relative to the geometric mean of the housekeeping genes GAPDH and ACTB (Vandesompele et al., 2002 (link)). Primers were designed using NCBI primer-BLAST. Primers sequences were as follows: C4A forward, 5′-GGCTCACAGCCTTTGTGTTG-3′; C4A reverse, 5′-CCCTGCATGCTCCTGTCTAA-3′;STAT1 forward, 5′-GCCAAAGGAAGCACCAGAGCCAAT-3′;STAT1 reverse, 5′-AGGAGACATGGGGAGCAGGTTGT-3′; IRF-1 forward, 5′-ATGAGACCCTGGCTAGAG-3′, IRF-1 reverse, 5′-AAGCATCCGGTACACTCG-3′; GAPDH forward, 5′-CGAGATCCCTCCAAAATCAA-3′; GAPDH reverse, 5′-TTCACACCCATGACGAACAT-3′; ACTB forward, 5′-TGAAGGTAGTTTCGTGGATGC-3′; ACTB reverse, 5′-TCCCTGGAGAAGAGCTACGA-3′. The C4A PCR product was sent to University of Illinois at Chicago DNA Services for sequencing to confirm specificity for C4A mRNA.

Melbourne J.K., Rosen C., Feiner B, & Sharma R.P. (2018). C4A mRNA expression in PBMCs predicts the presence and severity of delusions in schizophrenia and bipolar disorder with psychosis. Schizophrenia research, 197, 321-327.

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Quantitative PCR Analysis of Leukemia Cells

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For the real-time fluorescence quantitative PCR [reverse transcription (RT)–qPCR], the KG-1a cells, HL-60 cells, KG-1 cells, and THP-1 cells in good growth condition were selected and inoculated into the six-well-plate with 1 × 10⁶ cells per well. The total RNA was extracted using a traditional method of TRIzol (Cwbio, China) according to the instructions of the manufacturer. Validation of RNA-seq on 10 genes was performed using the RT-qPCR. RT and qPCR reactions were performed using the PrimeScript RT Master Mix (Takara, Japan) and the TB Green Premix Ex Taq II (Takara, Japan). The qPCR reaction was performed using the PikoReal (Thermo Fisher Scientific, United States), and qPCR data were analyzed by the ΔΔCt method. Primers used for RT-qPCR are listed in Supplementary Table 1.

Li Y., Feng Y., Si X., Zhao C., Wang F, & Niu X. (2021). Genetic Expression Screening of Arsenic Trioxide-Induced Cytotoxicity in KG-1a Cells Based on Bioinformatics Technology. Frontiers in Genetics, 12, 654826.

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Quantitative Analysis of STAT3 Target Genes

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MDA-MB-468 cancer cells were treated with 0, 50, or 100 μM BENDA for 8 h. Total RNA was extracted with RNeasy kits (Qiagen, Valencia, CA, USA) and reverse-transcribed to cDNA with the One-Step SYBR PrimeScript RT-PCR Kit II (Takara bio, Shiga, Japan). PCR amplification was performed under the following conditions: 5 min at 42°C, 10 s at 95°C, 40 cycles of 30 s at 94°C, 30 s at 55°C and 30 s at 72°C, and a final 5-min extension at 72°C. The DNA sequences of the primers for the STAT3 downstream target genes used for RT-PCR analysis were c-myc (forward: 5’-TACCCTCTCA ACGACAGCAG-3’, reverse: 5’-TCTTGACATT CTCCTCGGTG-3’) [29 (link)], cyclin D1 (forward: 5’-GCTGGAGCCC GTGAAAAAGA-3’, reverse: 5’-CTCCGCCTCT GGCATTTTG-3’), survivin (forward: 5’-ACCAGGTGAG AAGTGAGGGA-3’, reverse: 5’-AACAGTAGAG GAGCCAGGGA-3’), Bcl-2 (forward: 5’-TCTTTGAGTT CGGTGGGGTC-3’, reverse: 5’-TGCATATTTG TTTGGGGCAGG-3’), and GAPDH (forward: 5’-TGATGACATC AAGAAGGTGG TGAAG-3’, reverse: 5’-TCCTTGGAGG CCATGTGGGC AT-3’) [30 (link)]. The mRNA expression levels of the STAT3 target genes in cells treated with BENDA or HP2 were evaluated by quantitative RT-PCR (PikoReal, Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s protocol. Primers were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Iwamoto K., Uehara Y., Inoue Y., Taguchi K., Muraoka D., Ogo N., Matsuno K, & Asai A. (2017). Inhibition of STAT3 by Anticancer Drug Bendamustine. PLoS ONE, 12(1), e0170709.

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