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Hsa mir200a

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Hsa-miR200a is a microRNA (miRNA) molecule that is part of the miR-200 family. miRNAs are small, non-coding RNA molecules that play a role in regulating gene expression. The miR-200 family is involved in various cellular processes, but a detailed description of the core function of Hsa-miR200a cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Tri reagent, by Merck Group (2 mentions) Actinomycin d, by Merck Group (2 mentions) Click it nascent rna capture kit, by Thermo Fisher Scientific (2 mentions) Hsa mir200c, by Thermo Fisher Scientific (2 mentions) Pri mir200a, by Thermo Fisher Scientific (2 mentions) Pri mir200b, by Thermo Fisher Scientific (2 mentions) Pri mir429, by Thermo Fisher Scientific (2 mentions) Pri mir200c, by Thermo Fisher Scientific (2 mentions) Hsa mir200b, by Thermo Fisher Scientific (2 mentions) Hsa mir429, by Thermo Fisher Scientific (2 mentions)

2 protocols using hsa mir200a

1

Quantification of miRNA Biogenesis and Stability

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RNA was isolated using TRI reagent (Sigma) and samples were reverse transcribed using qScript cDNA SuperMix (Quantas). Quantitative real-time PCR was performed on an Eppendorf Realplex2 (link) mastercycler using SYBR green (Quantas). Specific primers used are listed in Supplementary Table 2. For determination of DICER mRNA half life cells were treated with 5 μg/ml actinomycin D (Sigma) for the indicated times. De novo RNA synthesis was measured using the Click-iT Nascent RNA Capture Kit (Molecular Probes). Briefly, MCF7 and HMLER cells were pulse labeled with 0.2mM 5-ethynyl Uridine for 1 hour during aerobic conditions or 24 hrs hypoxia (0.2% O2) after which RNA was isolated as described above. Nascent RNA was captured using magnetic streptavidin beads, reverse transcribed and analyzed by quantitative real-time PCR. For determination of pri-miRNA and mature miRNA levels the following assays from Applied Biosystems were used: pri-miR200a (Hs03303376_pri), pri-miR200b (Hs03303027_pri), pri-miR429 (Hs03303727_pri), pri-miR200c (Hs03303157_pri), pri-miR141 (Hs03303157_pri), pri-miR210 (Hs03302948_pri), hsa-miR200a (000502), hsa-miR200b (002251), hsa-miR429 (001024), hsa-miR200c (002300), miR141 (002145), hsa-miR103 (000439), hsa-miR107 (000443), hsa-miR210 (000512), RNU44 (001094), RNU48 (001006).
van den Beucken T., Koch E., Chu K., Rupaimoole R., Prickaerts P., Adriaens M., Voncken J.W., Harris A.L., Buffa F.M., Haider S., Starmans M.H., Yao C.Q., Ivan M., Ivan C., Pecot C.V., Boutros P.C., Sood A.K., Koritzinsky M, & Wouters B.G. (2014). Hypoxia promotes stem cell phenotypes and poor prognosis through epigenetic regulation of DICER. Nature communications, 5, 5203.
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2

Quantification of miRNA Biogenesis and Stability

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNA was isolated using TRI reagent (Sigma) and samples were reverse transcribed using qScript cDNA SuperMix (Quantas). Quantitative real-time PCR was performed on an Eppendorf Realplex2 (link) mastercycler using SYBR green (Quantas). Specific primers used are listed in Supplementary Table 2. For determination of DICER mRNA half life cells were treated with 5 μg/ml actinomycin D (Sigma) for the indicated times. De novo RNA synthesis was measured using the Click-iT Nascent RNA Capture Kit (Molecular Probes). Briefly, MCF7 and HMLER cells were pulse labeled with 0.2mM 5-ethynyl Uridine for 1 hour during aerobic conditions or 24 hrs hypoxia (0.2% O2) after which RNA was isolated as described above. Nascent RNA was captured using magnetic streptavidin beads, reverse transcribed and analyzed by quantitative real-time PCR. For determination of pri-miRNA and mature miRNA levels the following assays from Applied Biosystems were used: pri-miR200a (Hs03303376_pri), pri-miR200b (Hs03303027_pri), pri-miR429 (Hs03303727_pri), pri-miR200c (Hs03303157_pri), pri-miR141 (Hs03303157_pri), pri-miR210 (Hs03302948_pri), hsa-miR200a (000502), hsa-miR200b (002251), hsa-miR429 (001024), hsa-miR200c (002300), miR141 (002145), hsa-miR103 (000439), hsa-miR107 (000443), hsa-miR210 (000512), RNU44 (001094), RNU48 (001006).
van den Beucken T., Koch E., Chu K., Rupaimoole R., Prickaerts P., Adriaens M., Voncken J.W., Harris A.L., Buffa F.M., Haider S., Starmans M.H., Yao C.Q., Ivan M., Ivan C., Pecot C.V., Boutros P.C., Sood A.K., Koritzinsky M, & Wouters B.G. (2014). Hypoxia promotes stem cell phenotypes and poor prognosis through epigenetic regulation of DICER. Nature communications, 5, 5203.
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