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Mir x mirna first strand cdna synthesis kit

Manufactured by Takara Bio
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Sourced in Japan, United States

The Mir-X™ miRNA First Strand cDNA Synthesis Kit is a tool used for the reverse transcription of mature microRNA (miRNA) molecules into complementary DNA (cDNA) for subsequent analysis or downstream applications.

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Lab products found in correlation

Quant it ribogreen rna reagent and kit, by Thermo Fisher Scientific (3 mentions) Steponeplus system, by Thermo Fisher Scientific (2 mentions) Revertra ace rt kit, by Toyobo (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Kod plus mutagenesis kit, by Toyobo (1 mentions) Pegfp c1, by BD (1 mentions) Kapa sybr fast qpcr kit, by Roche (1 mentions)

6 protocols using mir x mirna first strand cdna synthesis kit

1

Quantifying RNA Concentration and Synthesizing cDNA

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The concentration of isolated RNA was measured using the Quant-IT RiboGreen RNA Reagent and Kit (Invitrogen, Grand Island, NY, USA). The cDNA was synthesized using the Mir-X™ miRNA First Strand cDNA Synthesis Kit (TAKARA BIO, Otsu, Japan) according to the manufacturer's protocol.
Byun Y.J., Piao X.M., Jeong P., Kang H.W., Seo S.P., Moon S.K., Lee J.Y., Choi Y.H., Lee H.Y., Kim W.T., Lee S.C., Cha E.J., Yun S.J, & Kim W.J. (2021). Urinary microRNA-1913 to microRNA-3659 expression ratio as a non-invasive diagnostic biomarker for prostate cancer. Investigative and Clinical Urology, 62(3), 340-348.
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2

Quantifying FFPE RNA for miRNA analysis

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The concentration of RNA isolated from FFPE tissues was measured using the Quant-IT RiboGreen RNA Reagent and Kit (Invitrogen, Grand Island, NY, USA). The Mir-X™ miRNA First Strand cDNA Synthesis Kit (TAKARA BIO, Otsu, Japan) was used to synthesize miRNA-specific cDNA according to the manufacturer’s protocol.
Byun Y.J., Kang H.W., Piao X.M., Zheng C.M., Moon S.K., Choi Y.H., Kim W.T., Lee S.C., Yun S.J, & Kim W.J. (2021). Expression of hsv1-miR-H18 and hsv2-miR-H9 as a field defect marker for detecting prostate cancer. Prostate International, 10(1), 1-6.
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3

RNA Quantification and cDNA Synthesis

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The concentration of isolated RNA was measured using the Quant-IT RiboGreen RNA Reagent and Kit (Invitrogen). The cDNA was synthesized using the Mir-X™ miRNA First Strand cDNA Synthesis Kit (Clontech, TAKARA Bio Inc., Otsu, Japan) according to the manufacturer’s protocol.
Kang H.W., Byun Y.J., Moon S.M., Kim K., Piao X.M., Zheng C.M., Moon S.K., Choi Y.H., Kim W.T., Kim Y.J., Lee S.C., Yun S.J, & Kim W.J. (2022). Urinary hsv2-miR-H9 to hsa-miR-3659 ratio is an effective marker for discriminating prostate cancer from benign prostate hyperplasia in patients within the prostate-specific antigen grey zone. Investigative and Clinical Urology, 63(2), 238-244.
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4

Gene Expression Analysis via RT-qPCR

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Total RNAs were isolated from cell lines or sphere using TRIzol® Reagent (Life Technologies, USA). For the analysis of mRNA, complementary DNA (cDNA) was synthesized by reverse transcription using a ReverTra Ace® RT Kit (Toyobo, Japan). For miRNA analysis, cDNA was prepared using the MiR-X™ miRNA First-Strand cDNA synthesis kit (Clontech, USA) according to the manufacturer’s instructions. The relative abundance of each transcript was assessed by real-time quantitative polymerase chain reaction (RT-qPCR) using the Bioline SYBR Fast qPCR kit (Bioline, UK) and specific primer sets on the StepOne Plus™ system (Applied Biosystems, USA).
Kang H., Kim C., Ji E., Ahn S., Jung M., Hong Y., Kim W, & Lee E.K. (2019). The MicroRNA-551a/MEF2C Axis Regulates the Survival and Sphere Formation of Cancer Cells in Response to 5-Fluorouracil. Molecules and Cells, 42(2), 175-182.
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5

Quantifying miRNA Levels and Regulation

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The MiR-X miRNA First-Strand cDNA synthesis kit (Clonetech, Mountain View, CA, USA) was used to construct cDNA from total RNA purified by TRIzol (Invitrogen). RT-qPCR was performed using the specific primers listed in Supporting Information to determine the miRNA levels. For miRNA reporter gene assays, the 3′UTR of human SETD1A (5948–6,271 bp) was inserted into pEGFP-C1 (BD Bioscience) to prepare EGFP reporter constructs, as described previously.25 (link) A mutant reporter construct was generated using a KOD plus mutagenesis kit (Toyobo, Osaka, Japan). The reporter construct or the miRNA was transfected using Lipofectamine 2000 (Invitrogen). Tissue sample are described in the Supporting Information.
Li Jin M., Kim Y.W., Jin H.L., Kang H., Lee E.K., Stallcup M.R, & Jeong K.W. (2018). Aberrant expression of SETD1A promotes survival and migration of estrogen receptor α-positive breast cancer cells. International journal of cancer, 143(11), 2871-2883.
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6

Quantification of mRNA and miRNA Transcripts

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Total RNAs were isolated from cell lines using Trizol reagent (Invitrogen, CA). For the analysis of mRNA, complementary DNA (cDNA) was synthesized by reverse transcription using a ReverTra Ace® RT Kit (Toyobo, Japan). For miRNA analysis, cDNA was prepared using the MiR-X™ miRNA First-Strand cDNA synthesis kit (Clonetech, CA) according to the manufacturer’s instructions. The relative abundance of each transcript was assessed by real-time quantitative polymerase chain reaction (RT-qPCR) using the Kapa SYBR Fast qPCR kit (Kapa Biosystems, MA) and specific primer sets on the StepOne Plus™ system (Applied Biosystems, CA). Primer sequences are listed in Table 1.
Kang H., Rho J.G., Kim C., Tak H., Lee H., Ji E., Ahn S., Shin A.R., Cho H.I., Huh Y.H., Song W.K., Kim W, & Lee E.K. (2017). The miR-24-3p/p130Cas: a novel axis regulating the migration and invasion of cancer cells. Scientific Reports, 7, 44847.
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