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Specific taqman primers

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Specific TaqMan primers are oligonucleotide sequences designed to amplify and detect specific target DNA or RNA sequences in a real-time PCR assay. They are used as part of the TaqMan probe-based detection system to enable sensitive and specific quantification of target nucleic acids.

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Lab products found in correlation

Mirneasy kit, by Qiagen (5 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (5 mentions) Specific taqman probes, by Thermo Fisher Scientific (4 mentions) Fast sybr green master mix, by Thermo Fisher Scientific (4 mentions) High capacity cdna synthesis kit, by Thermo Fisher Scientific (4 mentions) Superscript 2, by Thermo Fisher Scientific (2 mentions) Pcr master mix, by Thermo Fisher Scientific (2 mentions) Rneasy micro kit, by Qiagen (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) 2100 bioanalyzer, by Agilent Technologies (1 mentions) Abi viia 7, by Thermo Fisher Scientific (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Superscript 3 first strand synthesis kit, by Thermo Fisher Scientific (1 mentions) 7900ht fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Taqman fast advanced master mix, by Thermo Fisher Scientific (1 mentions) Transcriptor first strand cdna synthesis kit, by Roche (1 mentions) Taqman gene expression master mix, by Thermo Fisher Scientific (1 mentions) Omniscript rt kit, by Qiagen (1 mentions) High capacity rna to cdna kit, by Thermo Fisher Scientific (1 mentions)

14 protocols using specific taqman primers

1

Time-course Analysis of IAV-induced miRNA

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A time course study (0–8 h) on putative miRNA expression in cells exposed to IAV was carried out as described previously [7 (link)]. Total RNA, including miRNA, was isolated from HBEpCs infected with IAV H1N1 and H9N1 (1P10) or mock-infected cells using an miRNeasy kit (Qiagen, Germantown, MD, USA) following the protocol of the supplier. The quality of the total RNA was verified by an Agilent 2100 Bioanalyzer (Santa Clara, CA, USA) profile following the procedure as suggested by the manufacturer. RT-PCRs were performed to determine the expression of selected miRNAs (put-miR-31, 34, 35) on cDNA synthesized from total RNAs, including miRNAs, isolated from HBEpCs. Specific TaqMan primers were synthesized (Thermo Fisher Scientific, Carlsbad, CA, USA) and used with the TaqMan assay as described earlier [6 (link)]. Fold changes of miRNA were calculated using the ddCt method, where Ct is the threshold cycle to detect fluorescence. The data were normalized to control, and the fold change of miRNAs was calculated against control miRNA (U6 snRNA).
Othumpangat S., Beezhold D.H., Umbright C.M, & Noti J.D. (2021). Influenza Virus-Induced Novel miRNAs Regulate the STAT Pathway. Viruses, 13(6), 967.
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2

Quantifying Gene Expression in GLUTag Cells

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Extraction of RNA from siRNA-treated GLUTag cells was performed using an RNeasy Micro Kit (Qiagen). RNA (500 ng) was reverse-transcribed using SuperScript II (Thermo Fisher Scientific), and the resulting cDNA template was mixed with PCR Master Mix (Thermo Fisher Scientific), RNase-free water and specific TaqMan primers (Thermo Fisher Scientific). All experiments were performed on isolated cDNA samples from three independent transfection experiments. In all cases, expression was compared with that of β-actin measured from the same sample in parallel on the same plate, giving a Ct difference (ΔCt) for β-actin (Actb) minus the test gene. Expression is calculated as 2ΔCt . The following primer pairs were used (Thermo Fisher Scientific): Actb, Mm02619580; Lpl, Mm00434764_m1; Ffar1, Mm00809442_s1.
Psichas A., Larraufie P.F., Goldspink D.A., Gribble F.M, & Reimann F. (2017). Chylomicrons stimulate incretin secretion in mouse and human cells. Diabetologia, 60(12), 2475-2485.
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3

Quantitative RT-PCR Analysis of Gene Expression

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Extraction of RNA from siRNA-treated GLUTag cells was performed using an RNeasy Micro Kit (Qiagen). RNA (500 ng) was reverse-transcribed using SuperScript II (Thermo Fisher Scientific), and the resulting cDNA template was mixed with PCR Master Mix (Thermo Fisher Scientific), RNase-free water and specific TaqMan primers (Thermo Fisher Scientific). All experiments were performed on isolated cDNA samples from three independent transfection experiments. In all cases, expression was compared with that of β-actin measured from the same sample in parallel on the same plate, giving a Ct difference (ΔCt) for β-actin (Actb) minus the test gene. Expression is calculated as 2ΔCt. The following primer pairs were used (Thermo Fisher Scientific): Actb, Mm02619580; Lpl, Mm00434764_m1; Ffar1, Mm00809442_s1.
Psichas A., Larraufie P.F., Goldspink D.A., Gribble F.M, & Reimann F. (2017). Chylomicrons stimulate incretin secretion in mouse and human cells. Diabetologia, 60(12), 2475-2485.
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4

Quantitative HPV-16 Gene Expression

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Specific Taqman primers (Life Technologies) have been designed for the amplification of a region spanning HPV-16 E6 and E7 genes (Table 1D). Taqman GAPDH primers were used as endogenous control (Catalogue number 4333764 T, Life Technologies). cDNA samples were synthesized from DNAse-treated RNA as detailed above and were run in duplicate in qRT-PCR mix containing at least 25 ng cDNA, 1x Taqman Universal PCR master mix (Life Technologies) and 1x Taqman primer mix (Life Technologies). qRT-PCR was run on ABI Viia7 (Life Technologies) with the following conditions: Hold 50 °C for 2 mins; 10 mins of denaturation at 95 °C.; 40 cycles of: 95 °C (15 s), 60 °C (60s). Standard curves for the HPV-16 viral RNA copy number were developed using serial dilutions of cDNA synthesized from Caski-derived RNA. Standard curves were similarly developed for the GAPDH housekeeping gene. Viral RNA load was calculated from these standard curves as for DNA copy number, using similarly defined criteria for maximum cycle number to categorize a sample as HPV RNA positive.
Chai R.C., Lim Y., Frazer I.H., Wan Y., Perry C., Jones L., Lambie D, & Punyadeera C. (2016). A pilot study to compare the detection of HPV-16 biomarkers in salivary oral rinses with tumour p16INK4a expression in head and neck squamous cell carcinoma patients. BMC Cancer, 16, 178.
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5

Quantification of lncRNA and mRNA Levels

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Total RNA was isolated from cells using RNeasy Mini Kit (Qiagen, Valencia, CA). Reverse transcription of lncRNAs and mRNAs was performed using Superscript III First Strand Synthesis kit (Invitrogen, Carlsbad, CA) with random primers. qRT-PCR was carried out in the 7900HT Fast Real-Time PCR System (Applied Biosystems, Carlsbad, CA) using TaqMan Gene Expression Assay and gene-specific TaqMan primers (Life Technologies, Carlsbad, CA). Relative quantity of expression was calculated with the ΔΔCt method using rRNA 18S as an internal control. Samples were analyzed in quadruplicate.
Han Y.J., Boatman S.M., Zhang J., Du X.C., Yeh A.C., Zheng Y., Mueller J, & Olopade O.I. (2018). LncRNA BLAT1 is Upregulated in Basal-like Breast Cancer through Epigenetic Modifications. Scientific Reports, 8, 15572.
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6

Quantitative RT-PCR Analysis of Gene Expression

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Total RNA was prepared using the QIAGEN RNeasy Mini Kit, and cDNA was prepared with the Transcriptor First Strand cDNA synthesis kit (Roche), both following the manufacturer’s protocol. QPCR reactions were carried using TaqMan™ Fast Advanced Master Mix (Applied Biosystems) with commercially available specific Taqman primers (Life Technologies). Samples were run in duplicates and the QPCR assays were run on the Applied Biosystems real-time QPCR machine. Mouse samples used β-tubulin (tubb5) as the reference gene, and human samples used GAPDH.
Xie M.M., Amet T., Liu H., Yu Q, & Dent A.L. (2016). AMP Kinase Promotes Bcl6 Expression in both Mouse and Human T cells. Molecular immunology, 81, 67-75.
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7

Comprehensive RNA Analysis Pipeline

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Total RNA was isolated from eWAT and liver using the miRNeasy kit (Qiagen, Hilden, Germany) and assessed for quality (Agilent Bioanalyzer). Samples with RIN < 7 were excluded (n = 1 for WT-HFD eWAT and liver). For gene expression analysis, cDNA was synthesized from 200 ng of total RNA with the High-Capacity cDNA Synthesis kit (Applied Biosystems, Foster City, CA). qRT-PCR was performed using the 2x Fast SYBR Green Master Mix (Applied Biosystems; primer sequences in Supplementary Table 1). miRNAs were reverse-transcribed with specific TaqMan primers (Applied Biosystems), and subsequently measured by real-time PCR with the TaqMan universal PCR master mix and specific TaqMan probes (Applied Biosystems).
Shah A.P., Johnson M.D., Fu X., Boersma G.J., Shah M., Wolfgang M.J., Tamashiro K.L, & Baraban J.M. (2019). Deletion of translin (Tsn) induces robust adiposity and hepatic steatosis without impairing glucose tolerance. International journal of obesity (2005), 44(1), 254-266.
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8

Comprehensive RNA Analysis Pipeline

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Total RNA was isolated from eWAT and liver using the miRNeasy kit (Qiagen, Hilden, Germany) and assessed for quality (Agilent Bioanalyzer). Samples with RIN < 7 were excluded (n = 1 for WT-HFD eWAT and liver). For gene expression analysis, cDNA was synthesized from 200 ng of total RNA with the High-Capacity cDNA Synthesis kit (Applied Biosystems, Foster City, CA). qRT-PCR was performed using the 2x Fast SYBR Green Master Mix (Applied Biosystems; primer sequences in Supplementary Table 1). miRNAs were reverse-transcribed with specific TaqMan primers (Applied Biosystems), and subsequently measured by real-time PCR with the TaqMan universal PCR master mix and specific TaqMan probes (Applied Biosystems).
Shah A.P., Johnson M.D., Fu X., Boersma G.J., Shah M., Wolfgang M.J., Tamashiro K.L, & Baraban J.M. (2019). Deletion of translin (Tsn) induces robust adiposity and hepatic steatosis without impairing glucose tolerance. International journal of obesity (2005), 44(1), 254-266.
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9

Quantitative Real-Time PCR for Gene Expression

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Total RNA (2 μg) was reverse transcribed to cDNA using the Omniscript RT kit (Qiagen, Hilden, Germany) according to the manufacturer’s protocol. The cDNA served as a template for PCR amplification using the TaqMan Gene Expression Master Mix (Applied Biosystems, Foster City, CA, USA) and specific TaqMan primers (Applied Biosystems; Table 1) under the following conditions: 95°C for 10 min, followed by 40 cycles at 95°C for 15 s, and 60°C for 1 min. For assessment of relative quantities, the gene amounts were first normalized to a housekeeping gene, glyceraldehyde 3 phosphate dehydrogenase, followed by normalization to NG. Data are representative of three independent experiments.
Nam B., Kim S.A., Nam W., Jeung W.H., Park S.D., Lee J.L., Sim J.H, & Jang S.S. (2019). Lactobacillus plantarum HY7714 Restores TNF-α Induced Defects on Tight Junctions. Preventive Nutrition and Food Science, 24(1), 64-69.
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10

Comparative Gene Expression Analysis

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Total RNA was isolated from the cerebellum, eWAT and liver using the miRNeasy kit (Qiagen, Hilden, Germany). For gene expression analysis, cDNA was synthesized from 200 ng of total RNA with the High-Capacity cDNA Synthesis kit (Applied Biosystems, Foster City, CA). qRT-PCR was performed using the 2x Fast SYBR Green Master Mix (Applied Biosystems). MicroRNAs were reverse-transcribed with specific Taqman primers (Applied Biosystems) and subsequently measured by real-time PCR with the Taqman universal PCR master mix and specific Taqman probes (Applied Biosystems). The sequences of primers are shown in Supplementary Table 2.
Fu X., Shah A.P., Li Z., Li M., Tamashiro K.L, & Baraban J.M. (2020). Genetic inactivation of the translin/trax microRNA-degrading enzyme phenocopies the robust adiposity induced by Translin (Tsn) deletion. Molecular Metabolism, 40, 101013.
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