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Human mitochondrial dna mtdna monitoring primer set

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The Human Mitochondrial DNA (mtDNA) Monitoring Primer Set is a collection of oligonucleotide primers designed for the amplification and detection of human mitochondrial DNA sequences. The primer set is intended for use in various molecular biology applications that require the monitoring or analysis of human mitochondrial DNA.

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Lab products found in correlation

Sybr greener qpcr supermix, by Thermo Fisher Scientific (2 mentions) Abi prism 7900ht sequence detection, by Thermo Fisher Scientific (2 mentions) Qiaamp dna mini kit, by Qiagen (2 mentions) Power sybr green pcr master mix, by Thermo Fisher Scientific (2 mentions) Dneasy blood tissue kit, by Qiagen (1 mentions) Dnazol reagent, by Thermo Fisher Scientific (1 mentions) 7500 fast dx, by Thermo Fisher Scientific (1 mentions) Ezup column animal genomic dna purification kit, by Sangon (1 mentions) Purelink genomic dna mini kit, by Thermo Fisher Scientific (1 mentions) Fast real time pcr system, by Thermo Fisher Scientific (1 mentions) Kapa sybr fast qpcr kit master mix, by Roche (1 mentions) Genelute mammalian genomic dna miniprep kit, by Merck Group (1 mentions) Thermal cycler dice real time system single, by Takara Bio (1 mentions) Dneasy blood and tissue kit, by Qiagen (1 mentions)

Spelling variants of this product

Human Mitochondrial DNA Monitoring Primer Set (8 citations) MtDNA) Monitoring Primer Set (6 citations) Human mtDNA Monitoring Primer Set (3 citations) Primer sets (2 citations) Clontech Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (2 citations)

17 protocols using human mitochondrial dna mtdna monitoring primer set

1

Quantitative Analysis of Mitochondrial DNA

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DNA from HEK293A cells was extracted using the QIAamp DNA Mini Kit (Qiagen) according to the manufacturer’s instructions. The SyBR GreenER qPCR SuperMix (Invitrogen) was used for quantitative PCR with an ABI PRISM 7900HT sequence detection system (Applied Biosystems). The Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara) was used for amplification of mtDNA and nuclear DNA (nDNA), and data analysis was performed according to the manufacturer’s instructions.
Murakawa T., Yamaguchi O., Hashimoto A., Hikoso S., Takeda T., Oka T., Yasui H., Ueda H., Akazawa Y., Nakayama H., Taneike M., Misaka T., Omiya S., Shah A.M., Yamamoto A., Nishida K., Ohsumi Y., Okamoto K., Sakata Y, & Otsu K. (2015). Bcl-2-like protein 13 is a mammalian Atg32 homologue that mediates mitophagy and mitochondrial fragmentation. Nature Communications, 6, 7527.
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2

Quantifying Mitochondrial DNA Levels

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At the end of the experiment, total DNA was extracted from cells using a DNeasy Blood & Tissue Kit (Qiagen, Hilden, Germany). The ratio of mitochondrial mtDNA to nuclear DNA (mtDNA:nuDNA) was determined by quantitative PCR (qPCR) using a Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (TaKaRa, Kusatsu, Japan), in accordance with the manufacturer’s instructions. The kits included primers for ND1 and ND5 as the mtDNA targets and SLCO2B1 and SERPINA1 as the nuDNA targets. In brief, qPCR analysis was performed using the primer sets of ND1, ND5, SLCO2B1 and SERPINA1, and the threshold cycle (Ct) values were determined. Next, ΔCt1 (Ct value of SLCO2B1 − Ct value of ND1) and ΔCt2 (Ct value of SERPINA1 − Ct value of ND5) were calculated. Finally, the mean copy number of mtDNA was determined by calculating the mean value of 2ΔCt1 and 2ΔCt2.
Okuyama K., Kitajima Y., Egawa N., Kitagawa H., Ito K., Aishima S., Yanagihara K., Tanaka T, & Noshiro H. (2019). Mieap-induced accumulation of lysosomes within mitochondria (MALM) regulates gastric cancer cell invasion under hypoxia by suppressing reactive oxygen species accumulation. Scientific Reports, 9, 2822.
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3

Mitochondrial DNA Extraction and Quantification

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Mitochondrial DNA was extracted using a kit (Takara, Siga, Japan) in accordance with the manufacturer’s instructions. DNA samples were analyzed with a real-time PCR system. The primers used were the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara, Siga, Japan).
Araki C., Takemoto D., Kitagawa Y., Tateishi N., Rogi T., Izumo T., Kawamoto S., Shibata H., Hara E, & Nakai M. (2023). Sesamin Metabolites Suppress the Induction of Cellular Senescence. Nutrients, 15(7), 1627.
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4

Quantifying Mitochondrial DNA by PCR

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Mitochondrial DNA was quantified by real-time PCR using the human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara) according to the manufacturer’s guidelines. Primer sets for ND1 and ND5 were used for the detection of mtDNA and those of SLCO2B1 and SERPINA1were used for the detection of nuclear DNA.
Alharthi J., Bayoumi A., Thabet K., Pan Z., Gloss B.S., Latchoumanin O., Lundberg M., Twine N.A., McLeod D., Alenizi S., Adams L.A., Weltman M., Berg T., Liddle C., George J, & Eslam M. (2022). A metabolic associated fatty liver disease risk variant in MBOAT7 regulates toll like receptor induced outcomes. Nature Communications, 13, 7430.
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5

Quantification of Mitochondrial DNA

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Genomic DNA (gDNA) was extracted from HCT-116p53+/+ and HCT-116p53−/− cells by using DNAzol® Reagent (Thermo Scientific) and quantified using the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara Bio, Mountain View, CA, USA) according to manufacturer’s protocol. Primer sets for ND1 and ND5 were used for the detection of mtDNA, and the primer sets SLCO2B1 and SERPINA1were used for the detection of nuclear DNA. The quantitation of mtDNA copy number was performed using a 7500 FastDX (Applied Biosystems, MA, USA) and the power SYBR green PCR master mix (Applied Biosystem, Warrington WA1 4SR, UK).
De Vitto H., Ryu J., Calderon-Aparicio A., Monts J., Dey R., Chakraborty A., Lee M.H., Bode A.M, & Dong Z. (2020). Estrogen-related receptor alpha directly binds to p53 and cooperatively controls colon cancer growth through the regulation of mitochondrial biogenesis and function. Cancer & Metabolism, 8, 28.
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6

Quantifying Mitochondrial DNA Levels

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qRT-PCR analyses were performed to determine the mtDNA copy number in samples using the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara) in duplicates. Primers specific for SLCO2B1 and SERPINA1 were used for the determination of nuclear DNA (nDNA), and two primers (ND1, NADH dehydrogenase subunit 1, and ND5) were used for the detection of mtDNA. qRT-PCR was performed, and the mtDNA copy number was calculated according to the manufacturer’s protocol.
Ledrhem M., Nakamura M., Obitsu M., Hirae K., Kameyama J., Bouamama H., Gadhi C, & Katakura Y. (2022). Essential Oils Derived from Cistus Species Activate Mitochondria by Inducing SIRT1 Expression in Human Keratinocytes, Leading to Senescence Inhibition. Molecules, 27(7), 2053.
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7

Quantifying Human Mitochondrial DNA

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A total of 500 cells were sorted into Eppendorf tubes and lysed by flash freezing of pellets at −80°C. DNA was extracted with PicoPure DNA Extraction Kit, and Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (7246; Takara) was used to quantify the relative number of copies of human mtDNA by real-time PCR, with genomic DNA (gDNA) as the standard for normalization.
Docherty F.M., Riemondy K.A., Castro-Gutierrez R., Dwulet J.M., Shilleh A.H., Hansen M.S., Williams S.P., Armitage L.H., Santostefano K.E., Wallet M.A., Mathews C.E., Triolo T.M., Benninger R.K, & Russ H.A. (2021). ENTPD3 Marks Mature Stem Cell–Derived β-Cells Formed by Self-Aggregation In Vitro. Diabetes, 70(11), 2554-2567.
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8

Quantifying Mitochondrial DNA Levels

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Mitochondrial DNA copy number was measured using Human Mitochondrial DNA (mtDNA) monitoring Primer Set (TaKaRa) according to the manufacturer’s instructions.
Paul B.T., Tesfay L., Winkler C.R., Torti F.M, & Torti S.V. (2019). Sideroflexin 4 affects Fe-S cluster biogenesis, iron metabolism, mitochondrial respiration and heme biosynthetic enzymes. Scientific Reports, 9, 19634.
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9

Quantifying Mitochondrial DNA Content

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mtDNA content was measured by real-time polymerase chain reaction (RT-PCR) with the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara Japan). Briefly, we quantified mtDNA transcripts (NADH dehydrogenase subunit 1 (ND1) and NADH dehydrogenase subunit 5(ND5)), and normalized these to nuclear DNA transcripts (Solute Carrier 2B1(SLCO2B1) and Serpin peptidase inhibitor, clade A, member 1 (SERPINA1)).
Fujisawa K., Terai S., Takami T., Yamamoto N., Yamasaki T., Matsumoto T., Yamaguchi K., Owada Y., Nishina H., Noma T, & Sakaida I. (2016). Modulation of anti-cancer drug sensitivity through the regulation of mitochondrial activity by adenylate kinase 4. Journal of Experimental & Clinical Cancer Research : CR, 35, 48.
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10

Quantifying Mitochondrial DNA in Cells

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Mitochondrial DNA extracted from the cells was quantified using the Human Mitochondrial DNA (mtDNA) Monitoring Primer Set (Takara) according to manufacturer’s guidelines. Genomic DNA was extracted from MDA-MB-436 and SUM159PT cells which were transfected with MYC siRNA for 72 hours. Primer sets for ND1 and ND5 were used for the detection of mtDNA and those of SLCO2B1 and SERPINA1 were used for the detection of nuclear DNA.
Lee K.M., Giltnane J.M., Balko J.M., Schwarz L.J., Guerrero-Zotano A.L., Hutchinson K.E., Nixon M.J., Estrada M.V., Sánchez V., Sanders M.E., Lee T., Gómez H., Lluch A., Pérez-Fidalgo J.A., Wolf M.M., Andrejeva G., Rathmell J.C., Fesik S.W, & Arteaga C.L. (2017). MYC and MCL1 cooperatively promote chemotherapy-resistant breast cancer stem cells through regulation of mitochondrial oxidative phosphorylation. Cell metabolism, 26(4), 633-647.e7.
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