Mircury lna mirna pcr starter kit

Manufactured by Qiagen

Sourced in United States

The MiRCURY LNA miRNA PCR Starter Kit is a lab equipment product designed for the detection and quantification of microRNA (miRNA) expression using real-time PCR (Reverse Transcription Polymerase Chain Reaction) technology. The kit includes all the necessary reagents and components to perform miRNA profiling experiments.

Automatically generated - may contain errors

Lab products found in correlation

Mirneasy mini kit, by Qiagen (4 mentions) Mircury lna rt kit, by Qiagen (3 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (3 mentions) Sybr green reagent, by Qiagen (3 mentions) Primer express program, by Thermo Fisher Scientific (3 mentions) Abi prism 7900ht real time system, by Thermo Fisher Scientific (2 mentions) Mircury lna sybr green pcr kit, by Qiagen (2 mentions) Steponeplus, by Thermo Fisher Scientific (2 mentions) Mircury sybr green master mix, by Qiagen (1 mentions) Cfx96 connect real time pcr detection system, by Bio-Rad (1 mentions) Tri reagent, by Merck Group (1 mentions) Direct zol rna microprep kit, by Zymo Research (1 mentions) Cfx96 qpcr instrument, by Bio-Rad (1 mentions) Superscript 3 first strand synthesis system, by Thermo Fisher Scientific (1 mentions) Quantstudio software v1, by Thermo Fisher Scientific (1 mentions) Quantstudio 7 flex real time pcr system, by Thermo Fisher Scientific (1 mentions) Rnase free dnase 1, by Takara Bio (1 mentions) Reverse transcription master premix, by Elpis Biotech (1 mentions) Sybr premix ex taq, by Takara Bio (1 mentions) Rotor gene q, by Qiagen (1 mentions)

Spelling variants of this product

LNA miRCuRY (2 citations)

22 protocols using mircury lna mirna pcr starter kit

Quantification of miRNA Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA, including small RNA, was isolated from ECFCs using miRNeasy Mini Kit (QIAGEN, Hilden, DE) following the manufacturer’s instructions. For miRNA detection, 100 ng of total RNA was reverse transcribed by miRCURY LNA miRNA PCR Starter kit (QIAGEN) and detected using miRCURY SYBR Green Master Mix (QIAGEN) in CFX96 connect real-time PCR detection system (Bio-Rad, Hercules, CA, USA). miRNA specific primers (Table S1) were purchased from miRCURY LNA miRNA PCR Starter kit (QIAGEN). The expression levels of miRNAs were normalized to UniSp6, a spike-in control small nuclear RNA.

Tsai W.C., Chiang W.H., Wu C.H., Li Y.C., Campbell M., Huang P.H., Lin M.W., Lin C.H., Cheng S.M., Chang P.C, & Cheng C.C. (2020). miR-548aq-3p is a novel target of Far infrared radiation which predicts coronary artery disease endothelial colony forming cell responsiveness. Scientific Reports, 10, 6805.

+ Open protocol

+ Expand

Quantitative Analysis of Epithelial-Mesenchymal Transition

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed with TRI Reagent (Sigma-Aldrich). Total RNA was purified from cells using Direct-zol RNA Microprep Kit (Zymo Research, Irvine, CA, USA). cDNA synthesis from mRNA was carried out using a SuperScript III First-Strand Synthesis System (Invitrogen), and Takyo No Rox SYBR MasterMix dTTP Blue (Eurogentec, Fremont, CA, USA) was used for real-time PCR assay. For microRNA, the miRCURY LNA miRNA PCR Starter Kit (QIAGEN, Germantown, MD, USA) was used to construct cDNA and also real-time PCR assay. All qPCR assays were conducted in a CFX96 qPCR Instrument (Bio-Rad). Four independent repeats were performed. mRNA was normalized to human acidic ribosomal protein (hARP). miR-200c was normalized to miR-103a. The primers used for the PCR are:
CDH1: 5’-CAGGTCTCCTCTTGGCTCTG-3’ and 5’-GACCGGTGCAATCTTCAAAA-3’; CDH2: 5’-GTGCATGAAGGACAGCCTCT-3’ and 5’-AGCTTCTCACGGCATACACC-3’; VIM: 5’-CGAAAACACCCTGCAATCTT-3’ and 5’-CTGGATTTCCTCTTCGTGGA-3’; SNAI1: 5’-CTTCTCTAGGCCCTGGCTG-3’ and 5’-CATCTGAGTGGGTCTGGAGG-3’; SNAI2: 5’-TCGGACCCACACATTACCTT-3’ and 5’-TGACCTGTCTGCAAATGCTC-3’; TWIST1: 5’-GGACAAGCTGAGCAAGATTCA-3’ and 5’-CGGAGAAGGCGTAGCTGAG-3’; ZEB1: 5’-AGCAGTGAAAGAGAAGGGAATGC-3’ and 5’-GGTCCTCTTCAGGTGCCTCAG-3’; ZEB2: 5’-TTTCAGGGAGAATTGCTTGA-3’ and 5’-CACATGCATACATGCCACTC-3’; hARP: 5’-CACCATTGAAATCCTGAGTGATGT-3’ and 5’-TGACCAGCCCAAAGGAGAAG -3’.

Lin M., Ku A.T., Dong J., Yue F., Jiang W., Ibrahim A.A., Peng F., Creighton C.J., Nagi C., Gutierrez C., Rosen J.M., Zhang X.H., Hilsenbeck S.G., Chen X., Du Y.C., Huang S., Shi A., Fan Z, & Li Y. (2022). STAT5 confers lactogenic properties in breast tumorigenesis and restricts metastatic potential. Oncogene, 41(48), 5214-5222.

+ Open protocol

+ Expand

Quantification of miRNA Expression

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

To determine expression, RNA was extracted from both cell lines with the miRCURY LNA miRNA PCR Starter Kit (Qiagen). To carry out the quantitative real-time polymerase chain reaction (qRT-PCR), we followed the protocol described by Rama et al. [31 (link)] using miR-103a and miR-191 using miR-103a and miR-191 as housekeeping.

Rama A.R., Lara P., Mesas C., Quiñonero F., Vélez C., Melguizo C, & Prados J. (2022). Circular Sponge against miR-21 Enhances the Antitumor Activity of Doxorubicin against Breast Cancer Cells. International Journal of Molecular Sciences, 23(23), 14803.

+ Open protocol

+ Expand

Quantitative Analysis of Gene Expression

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA (125 ng) was used to generate cDNA using the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Carlsbad, CA) according to the manufacturer’s instructions. qPCR analysis was performed using the ABI Prism 7900HT Real-Time System instrument (Applied Biosystems) with SYBR Green reagent (Qiagen) for mRNA and as previously described (Farr et al., 2015 (link)). Data normalization was performed based on 5 reference genes (Actb, Gapdh, Polr2a, Rpl13a, Tuba1a) depending on their stability and threshold calculations are as previously described (I. Mödder et al., 2011 (link)). The oligonucleotide sequences for the genes measured in this study were designed using the Primer Express program (Applied Biosystems) and are available upon request. For the miRNA expression data, total RNA (30 ng) including the miRNA fraction was reverse transcribed using the miRCURY LNA RT Kit (Qiagen) as per manufacturer’s protocol. All the individual miRNA assays used in this study were purchased from Qiagen and used with the miRCURY LNA miRNA PCR Starter Kit (Qiagen) according to the manufacturer’s instructions. Data normalization was performed using the Let-7f and RNU6B (U6) small nucleolar RNA (snRNA) as specified in Results.

Kaur J., Saul D., Doolittle M.L., Rowsey J.L., Vos S.J., Farr J.N., Khosla S, & Monroe D.G. (2022). Identification of a suitable endogenous control miRNA in bone aging and senescence. Gene, 835, 146642.

+ Open protocol

+ Expand

Exosomal microRNA Quantification via qRT-PCR

Check if the same lab product or an alternative is used in the 5 most similar protocols

Custom 96-wells Pick-&-Mix microRNA PCR plates (Qiagen) were used to validate teach miRNA candidate. qRT-PCR was performed to quantify the expression levels of candidate exosomal miRNAs using an miRCURY LNA miRNA PCR Starter Kit (Qiagen, No. 339320) and a miRCURY LNA SYBR Green PCR Kit (Qiagen, No. 339347). In addition, we used the Applied Biosystems QuantStudio 7 Flex Real-Time PCR System in a total volume of 10 μL. qRT-PCR conditions were 95 °C for 2 min, followed by 40 cycles of 95 °C for 10 s and 56 °C for 1 min, followed by melting curve analysis. Synthetic UniSp3 was analyzed as interplate calibrator and qRT-PCR control. Amplification curves were evaluated using QuantStudio Software v1.3 (Thermo Fisher Scientific, Massachusetts, USA). Quantification cycles (Cq) > 35 cycles were censored at the minimum level observed for each miRNA. cel-miR-39-3p levels were stable across samples. Relative quantification was performed using the 2^−ΔCq method (ΔCq = Cq_miRNA-Cq_{cel-miR-39-3p}). Expression levels were log-transformed for statistical purposes [23 ].

Shin B., Lee J.Y., Im Y., Yoo H., Park J., Lee J.S., Lee K.Y, & Jeon K. (2023). Prognostic implication of downregulated exosomal miRNAs in patients with sepsis: a cross-sectional study with bioinformatics analysis. Journal of Intensive Care, 11, 35.

+ Open protocol

+ Expand

Quantitative Analysis of miR-18a-5p Expression

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total cellular RNA was extracted using TRIzol Reagent. Isolated RNAs were treated with RNase‐free DNase I (Takara Bio) and used for complementary DNA (cDNA) synthesis, which was conducted using the Reverse Transcription Master Premix (Elpis Biotech). qRT‐PCR was performed and analyzed using StepOnePlus (Applied Biosystems) with SYBR Premix Ex Taq (Takara Bio). The specific primer sequences are listed in Table 2. Glyceraldehyde 3‐phosphate dehydrogenase was used as a reference gene. Synthesis of cDNA from miR‐18a‐5p and quantification of miR‐18a‐5p transcript levels were performed using miRCURY LNA miRNA PCR Starter Kit (Qiagen) following the manufacturer's instructions. U6 was used as a control for normalization of miR‐18a‐5p.

Baek J., Shin H., Suk K, & Lee W. (2024). LINC01686 affects LPS‐induced cytokine expression via the miR‐18a‐5p/A20/STAT1 axis in THP‐1 cells. Immunity, Inflammation and Disease, 12(4), e1234.

+ Open protocol

+ Expand

miR-21-5p Expression Profiling

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cDNAs were produced from total RNA by reverse transcription using the miRCURY LNA miRNA PCR Starter Kit (Cat no. 339320, QIAGEN, Germantown, MD, USA). The miRNA PCR primer set, based on the SYBR Green miRCURY locked nucleic acids (LNA) detection system from QIAGEN, was used to profile the expression of miR-21-5p (Cat no. YP00204230, QIAGEN, Germantown, MD, USA). The expression level of UniSp6 (U6) was used as an endogenous control according to the manufacturer’s instructions.

Chen Y.C., Lai Y.S., Hsuuw Y.D, & Chang K.T. (2021). Withholding of M-CSF Supplement Reprograms Macrophages to M2-Like via Endogenous CSF-1 Activation. International Journal of Molecular Sciences, 22(7), 3532.

+ Open protocol

+ Expand

miR-155 Expression Analysis in RAW 264.7 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

RAW 264.7 cells (2 × 10⁵ cells/well) were seeded in 6-well plates and cultured to 60% confluency. The cells were incubated with the nanoparticles (100 nM of anti-miR-155) or 100 nM of anti-miR-155/Lipofectamine 2000 as the positive control for 6 h in a serum-free medium, then removing the nanoparticle solutions followed by stimulation with LPS (100 ng/mL) for 24 h. RT-PCR analysis was performed using a Rotor-Gene Q (QIAGEN) thermocycler with a miRCURY LNA miRNA PCR Starter Kit (QIAGEN). The universal reverse transcription was followed by real-time PCR amplification with the LNA-enhanced primers. MiR-155 expression was expressed relative to the internal control UniSp6.

Zhang C., Hang Y., Tang W., Sil D., Jensen-Smith H.C., Bennett R.G., McVicker B.L, & Oupický D. (2022). Dually Active Polycation/miRNA Nanoparticles for the Treatment of Fibrosis in Alcohol-Associated Liver Disease. Pharmaceutics, 14(3), 669.

+ Open protocol

+ Expand

Quantifying Dysregulated miRNAs in Lung Cancer

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA including miRNA was purified with the miRNeasy Mini Kit (#217004, Qiagen MD). The expression of miR-27a, miR-27b, miR-33a, miR-33b, miR-128, miR-125-3p, miR-371a-5p, miR-365, miR-425 (# E01007) and U6 (#E01008) was detected using Hairpin-it™ miRNAs qPCR Quantitation Kit (Genepharma, China). miR-125a-3p mimic (YM00471085-ADA, #339173 miRCURY LNA miRNA Mimic, Qiagen MD), miR-425 mimic (YM00471725-ADA, #339173 miRCURY LNA miRNA Mimic, Qiagen MD) miR-371-5p mimic (YM00472003-ADA, #339173 miRCURY LNA miRNA Mimic, Qiagen MD) and unrelated sequences miR-NC used as negative controls (YM00479902-ADA, #339173 miRCURY LNA miRNA Mimic, Qiagen MD) were provided by Qiagen. The expression of miR-125-3p, miR-371a-5p, and miR-425 in the mimic microRNAs transfected cells were detected by miRCURY LNA miRNA PCR Starter Kit (#339320, Qiagen MD). The expression of target miRNAs in the human lung tissues, nude mice tumors, or the treated and control cells was normalized using U6 or miR-NC controls and the fold change in the expression of each target gene was calculated.

Liu L.Z., Wang M., Xin Q., Wang B., Chen G.G, & Li M.Y. (2020). The permissive role of TCTP in PM2.5/NNK-induced epithelial–mesenchymal transition in lung cells. Journal of Translational Medicine, 18, 66.

+ Open protocol

+ Expand

Reverse Transcription and qPCR for miRNA Analysis

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

10ng of total RNA were reverse-transcribed and amplified using the miRCURY LNA miRNA PCR Starter Kit (Qiagen, Ref 339320. The kit includes a spike-in control primer set (UniSp6), UniSP6 RNA Spike-in-template, one candidate endogenous control primer set (miR-103a-3p) and two validated primer sets, which in our case were miR-297 (Qiagen YP00206079) and miR-574-5p (Qiagen YP02116206). As additional control micro-RNAs (miRNAs), we chose miR-25-3p (Qiagen YP00204361) and miR-331-3p (Qiagen YP00206046) because both are used as markers in BC analysis (27 (link), 28 (link)) and their expression is not modified by RA treatment in the BC cell line SKBR3 (11 (link)). Real-time PCR was performed with miRCURY LNA SYBR Green Master Mix (Qiagen) in 10μL total volume using the CFX 96 Real Time System (Bio-Rad). The expression of target miRNAs miR-297 and miR-574-5p was normalized against miR-25-3p, miR-103a-3p, miR-331-3p and UniSp6 using the 2^-ΔΔCt method. To validate the real-time system used for miRNA analysis, we measured the levels of UniSp6RNA, a control RNA provided with the Starter Kit, that was added before the reverse transcription in equal amount to all samples (see Supplementary material).

Piccolella M., Cristofani R., Tedesco B., Chierichetti M., Ferrari V., Casarotto E., Cozzi M., Crippa V., Rusmini P., Galbiati M., Poletti A, & Messi E. (2021). Retinoic Acid Downregulates HSPB8 Gene Expression in Human Breast Cancer Cells MCF-7. Frontiers in Oncology, 11, 652085.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!