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Mrq 3 primer

Manufactured by Takara Bio
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Sourced in Japan

The MRQ 3' Primer is a lab equipment product designed for use in molecular biology applications. It serves as a primer for reverse transcription and PCR amplification. The product provides a concise and functional description without any interpretations or extrapolations on its intended use.

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Lab products found in correlation

Mir x mirna first strand synthesis kit, by Takara Bio (4 mentions) Mirna mini kit, by Qiagen (2 mentions) Cfx96 real time qpcr instrument, by Bio-Rad (2 mentions) Primescript rt master mix, by Takara Bio (2 mentions) Tb green premix ex taq 2 reagent, by Takara Bio (1 mentions) U6 snrna specific primer, by Takara Bio (1 mentions) Primescript rt pcr kit, by Takara Bio (1 mentions) Lightcycler 96 instrument, by Roche (1 mentions) Sybr advantage qpcr premix, by Takara Bio (1 mentions) Sybr premix ex taq 2, by Takara Bio (1 mentions) Sybr premix ex taqtm 2, by Takara Bio (1 mentions) Mir xtm mirna first strand synthesis kit, by Takara Bio (1 mentions)

7 protocols using mrq 3 primer

1

Quantifying miR-28-5p and N4BP1 Expression

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The primer sequences of miR-28-5p were: forward 5'-CGC AAG GAG CTC ACA GTC TAT TGA G-3'; reverse: mRQ3'primer (Takara, Tokyo, Japan). The primer sequences of N4BP1were: forward 5'-CAC CTT CTG TTG CCT CTC CAA GTC-3'; reverse 5'-GGG TTC TGG CTG GTG TAA AC-3'.
Hu J.C., Zhu T.P., Gui Y.C., Tan Z.B., Wei R.Q., Hu B.L, & Xu J.W. (2020). miR-28-5p inhibits carcinogenesis in colon cancer cells and is necessary for erastin-induced ferroptosis. Translational Cancer Research, 9(4), 2931-2940.
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2

Quantifying Exosomal miRNAs in LUAD

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To explore the differentially expressed exosomal miRNAs in LUAD, the levels of six candidate miRNAs were examined using a relative quantification method. The extracted miRNA was reverse-transcribed with adding a poly (A) tail using the Mir-X miRNA First-Strand Synthesis Kit (TaKaRa Bio, Japan). The reaction mixture was incubated at 37°C for 60 min, and then 85°C for 5 min. qRT-PCR was performed using TB-Green Premix Ex Taq II Reagent (TaKaRa Bio, Japan). The mRQ 3′ Primer (TaKaRa Bio, Japan) was used as the reverse primer for each miRNA, and the sequences of miRNA-specific primers are provided in Table S1. The conditions of the amplification reaction were as follows: 95°C for 10 s, 40 cycles of 95°C for 5 s, and 60°C for 32 s. Each sample was analyzed in duplicate. Relative miRNA expression levels were calculated using the ΔCT method with U6 snRNA as a reference (CtmiRNA - CtU6).
Wu Y., Wei J., Zhang W., Xie M., Wang X, & Xu J. (2020). Serum Exosomal miR-1290 is a Potential Biomarker for Lung Adenocarcinoma. OncoTargets and therapy, 13, 7809-7818.
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3

Quantitative Analysis of Gene and miRNA Expression

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cDNA was synthesised from total RNA (500 ng) in a 10-μl reaction volume using a PrimeScript RT-PCR Kit (TaKaRa, USA). Thermal cycling conditions included an initial step at 98 °C for 30 s, followed by 40 cycles at 95 °C for 5 s and 60 °C for 20 s. The following primers were used: MMP-2 forward, 5′-GATAACCTGGATGCCGTCGTG-3′; MMP-2 reverse, 5′-CAGCCTAGCCAGTCGGATTTG-3′; TIMP-2 forward, 5′-TACCAGATGGGCTGCGAGTG-3′; TIMP-2 reverse, 5′-CTGTGACCCAGTCCATCCAGAG-3′; and GAPDH forward, 5′-CCATCACCATCTTCCAGGAG-3′; GAPDH reverse, 5′-CCTGCTTCACCACCTTCTTG-3′.
The relative expression levels of mRNA were analysed using the 2−ΔΔCt method. For miRNA analysis, cDNA was synthesised using a miR-X miRNA first-strand synthesis kit (TaKaRa, Japan). Thermal cycling conditions included an initial step at 98 °C for 30 s, followed by 40 cycles at 95 °C for 2 s and 63 °C for 5 s. The following primers were used: miR-130b primer, 5′-CAGTGCAATGATGAAAGGGCAT-3′; mRQ 3′ primer (TaKaRa); and U6 snRNA-specific primer (TaKaRa) as an internal control.
Hirono T., Jingushi K., Nagata T., Sato M., Minami K., Aoki M., Takeda A.H., Umehara T., Egawa H., Nakatsuji Y., Kitae K., Ueda Y., Hase H., Yamamoto M., Shinsato Y., Kawahara K., Furukawa T, & Tsujikawa K. (2019). MicroRNA-130b functions as an oncomiRNA in non-small cell lung cancer by targeting tissue inhibitor of metalloproteinase-2. Scientific Reports, 9, 6956.
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4

Mir-X miRNA First-Strand Synthesis Protocol

Cited 1 time
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The Mir-X miRNA First-Strand Synthesis kit (Clontech) was applied on total RNA (25 ng) or synthetic RNA (to obtain the miRNA standard curves as described for the miRCURY and miRPrimer strategies) according to the manufacturer’s instructions, in a final volume of 10 µL in the presence of mRQ enzyme. The reaction took place at 37 °C for 1 h. cDNA samples were diluted (40×) and 10 µL of PCR reaction including specific miRNA forward primer (the entire sequence of the miRNA isoform which included 4A) added to the 3′-end as previously described [14 (link)]; Table 1, the mRQ 3′ primer (Clontech) as reverse primer and 4.6 µL of diluted cDNA. Cycling conditions were identical to the ones used for miRPrimer2 strategy on the LightCycler® 96 Instrument (Roche; Switzerland).
Ferre A., Santiago L., Sánchez-Herrero J.F., López-Rodrigo O., Sánchez-Curbelo J., Sumoy L., Bassas L, & Larriba S. (2023). 3′IsomiR Species Composition Affects Reliable Quantification of miRNA/isomiR Variants by Poly(A) RT-qPCR: Impact on Small RNA-Seq Profiling Validation. International Journal of Molecular Sciences, 24(20), 15436.
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5

Real-time qPCR Validation of miRNA Expression

Cited 1 time
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The real-time qPCR validation of miRNA expression was performed using
Mir-X miRNA first-strand synthesis and quantification kit (Clontech), according
to the manufacturer's instructions. The real-time qPCR quantification of
specific miRNA sequences was performed on a 60-fold dilution of the cDNA samples
using the SYBR Advantage qPCR premix and mRQ 3’ Primer (Clontech). The
PCR reaction was carried out on a Stratagene Mx3005P instrument using the
conditions of 1-cycle denaturation at 95°C, 10 sec; 40-cycle
amplification at 95°C, 5 sec and 60°C, 20 sec; and dissociation
at 95°C, 60 sec; 55°C, 30 sec; 95°C, 30 sec. All
reactions were performed in triplicate and included with no template control for
each gene. U6 snRNA was included in each sample as an endogenous control to
determine the Ct for determination of the relative level using delta-delta
Ct (ddCt) calculation method [23 (link)], ΔCt=Ct
miRNA−Ct U6;
ΔΔCt=Ct test
group−ΔCtcontrol group; relative miRNA
expression=2−ΔΔCT.
The level of putative target mRNA was quantified by real-time RT-PCR as
described previously [28 (link)]. Expression
levels of the genes were normalized to 18S rRNA. Each sample
was compared in 3 independent RT-PCR amplifications.
Tang Y., Lu Q., Wei Y., Han L., Ji R., Li Q, & Lu Q. (2015). Mertk Deficiency Alters Expression of microRNAs in the Retinal Pigment Epithelium Cells. Metabolic brain disease, 30(4), 943-950.
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6

Analyzing miRNA and mRNA Expression

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Cartilage and cell-seeded scaffolds were ground in liquid nitrogen prior to RNA isolation. Total RNA from cells, cartilage samples, and cell-seeded scaffolds was extracted using a miRNA Mini Kit (Qiagen, Hilden, Germany) following the manufacturer's instructions.
Next, cDNA was synthesized from miRNA and mRNA using a Mir-X™ miRNA First-Strand Synthesis Kit (Clontech Laboratories, Inc., Mountain View, CA, USA) and a PrimeScript™ RT Master Mix (Takara, Shiga, Japan), respectively. qRT-PCR of target genes was performed using SYBR® Premix Ex Taq™ II (Takara) and a CFX96 real-time qPCR instrument (Bio-Rad, Hercules, CA, USA), according to the manufacturer's instructions. Transcript levels were normalized to that of the reference gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) for mRNA, the small U6 RNA for miRNA, or miR-39 for exosomal miRNA. The specific primers used for these analyses are shown in Supplementary Material. The mRQ 3′ Primer (Clontech) was used as the reverse primer for miRNA-193b-3p, and the miR-39 primer was supplied in the miRNeasy Serum/Plasma Kit. Gene expression was calculated using the 2-ΔΔCt method, and each experiment was performed in triplicate.
Meng F., Li Z., Zhang Z., Yang Z., Kang Y., Zhao X., Long D., Hu S., Gu M., He S., Wu P., Chang Z., He A, & Liao W. (2018). MicroRNA-193b-3p regulates chondrogenesis and chondrocyte metabolism by targeting HDAC3. Theranostics, 8(10), 2862-2883.
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7

Quantitative Analysis of miRNA and mRNA Levels

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With miRNA Mini Kit (Qiagen, Hilden, Germany), total RNA from cells and cartilage samples were extracted following manufacturer’s instructions. Next, cDNA was synthesized from miRNA and mRNA using a Mir-XTM miRNA First-Strand Synthesis Kit (Takara Bio Inc., Shiga, Japan) and a PrimeScriptTM RT Master Mix (Takara), respectively. RT-qPCR of target genes was performed with SYBR® Premix Ex TaqTM II (Takara) and a CFX96 real-time qPCR instrument (Bio-Rad, Hercules, CA, United States). Transcript levels were normalized to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) for mRNA and U6 RNA for miRNA, qPCR primer sequences can be found in Supplementary Table 1. The mRQ 3′ Primer (Clontech) was used as the reverse primer for miRNA-455-3p. Relative gene expression was calculated using the 2–ΔΔCt method, and each experiment was performed in triplicate.
Mao G., Kang Y., Lin R., Hu S., Zhang Z., Li H., Liao W, & Zhang Z. (2019). Long Non-coding RNA HOTTIP Promotes CCL3 Expression and Induces Cartilage Degradation by Sponging miR-455-3p. Frontiers in Cell and Developmental Biology, 7, 161.
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