mircury lna rt kit (Qiagen)

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miRCURY LNA RT Kit

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For optimal first strand cDNA synthesis with the miRCURY LNA miRNA PCR System Kit contents cDNA synthesis kit for reverse transcription of miRNA Benefits Optimized for use with miRCURY LNA miRNA PCR Assays and Panels Fast easy and reliable first strand synthesis Includes RNA spike in template for monitoring performance

Catalog Number:

339340

Price:

356

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miRCURY LNA RT Kit

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Qiagen mircury lna rt kit

mircury lna rt kit - by Bioz Stars, 2020-01

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Amplification:

Article Title: Bioinformatic analysis of long-lasting transcriptional and translational changes in the basolateral amygdala following acute stress
Article Snippet: QPCR validation For validation of smRNA-seq, a cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) following the manufacturer’s protocol and technical replicate samples that were sequenced. cDNA was diluted 1:80 for PCR. .. PCR cycle conditions were 95°C for ten minutes then 40 amplification cycles of 95°C for 10 seconds to denature and 60°C for 1 minute to anneal and elongate.

Article Title: MicroRNA-deficient mouse embryonic stem cells acquire a functional interferon response
Article Snippet: .. RNA was first converted to cDNA using miRCURY LNA RT kit (Qiagen). cDNA was diluted 1/25 for RT-qPCR using miRCURY LNA SYBR Green kit and amplified using mmu-mmiR-673–5p specific primers (Qiagen) and U6 as a loading control. .. Quantitative PCR was carried out on a Roche LC480 light cycler and analysed using the second derivative method.

Stable Transfection:

Article Title: Transcriptomic profiling of cell-free and vesicular microRNAs from matched arterial and venous sera
Article Snippet: To identify reference candidates, we evaluated the most stably expressed miRNAs from the small RNA-Seq dataset by geNorm [ ] and NormFinder [ ]. .. Four µl RNA were used as starting material for reverse transcription (miRCURY LNA RT Kit, Qiagen, Venlo, The Netherlands).

Synthesized:

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. PCR primers were synthesized by Sangon Biotech (Shanghai, China), and sequence information is exhibited in Table .

Article Title: Maternal circulating miRNAs that predict infant FASD outcomes influence placental maturation
Article Snippet: .. For miRNA qPCR assays, cDNA was synthesized from 200 ng of total RNA using the miRCURY LNA Universal RT cDNA synthesis kit (Cat No. 203301; Exiqon/Cat No. 339340; QIAGEN), and expression was assessed using miRCURY LNA SYBR Green (Cat No. 203401; Exiqon/Cat No. 339345; QIAGEN). .. For mRNA qPCR assays, cDNA was synthesized from 500 ng of total RNA using the qScript cDNA Synthesis kit (Cat No. 95047; Quanta/QIAGEN).

Article Title: Identification of stably expressed housekeeping miRNAs in endothelial cells and macrophages in an inflammatory setting
Article Snippet: .. Complementary DNA (cDNA) was synthesized by means of the miRCURY LNA RT Kit (QIAGEN, Hilden, Germany) according to standard protocol. .. Synthesized cDNA was used for both qRT-PCR and ddPCR miRNA expression quantification.

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. PCR primers were synthesized by Sangon Biotech (Shanghai, China), and sequence information is exhibited in Table .

Quantitative RT-PCR:

Article Title: Reprogramming of Small Noncoding RNA Populations in Peripheral Blood Reveals Host Biomarkers for Latent and Active Mycobacterium tuberculosis Infection
Article Snippet: RNA yield was measured with a NanoDrop S1000 spectrophotometer (Thermo Scientific), and 50 ng was then reverse transcribed using a miRCURY LNA RT kit (Qiagen). .. Quantitative real-time PCR (RT-qPCR) was performed using a SensiFAST SYBR kit (Bioline).

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Cellular microRNA detection with miRacles: microRNA- activated conditional looping of engineered switches
Article Snippet: .. qRT-PCR for miRNAs qRT-PCR for miRNAs was carried out by a miRCURY LNA RT kit (Qiagen) following the instructions of the manufacturer. .. miRNA detection In general, assays were performed by incubating the purified nanoswitches (~100 pM) with the target solution containing miRNAs.

Article Title: Down-regulation of long non-coding RNA MEG3 suppresses osteogenic differentiation of periodontal ligament stem cells (PDLSCs) through miR-27a-3p/IGF1 axis in periodontitis
Article Snippet: Paragraph title: Quantitative real-time PCR (qRT-PCR) ... Thereafter, about 2 μg of total RNA was reversely transcripted to cDNA using SuperScriptTM III Reverse Transcriptase Kit (Invitrogen) for lncRNA/mRNA reverse transcription and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) for miRNA reverse transcription.

Article Title: Identification of stably expressed housekeeping miRNAs in endothelial cells and macrophages in an inflammatory setting
Article Snippet: Complementary DNA (cDNA) was synthesized by means of the miRCURY LNA RT Kit (QIAGEN, Hilden, Germany) according to standard protocol. .. Synthesized cDNA was used for both qRT-PCR and ddPCR miRNA expression quantification.

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Transcriptomic profiling of cell-free and vesicular microRNAs from matched arterial and venous sera
Article Snippet: Paragraph title: RT-qPCR ... Four µl RNA were used as starting material for reverse transcription (miRCURY LNA RT Kit, Qiagen, Venlo, The Netherlands).

Article Title: Modified Polyadenylation-Based RT-qPCR Increases Selectivity of Amplification of 3′-MicroRNA Isoforms
Article Snippet: Paragraph title: Reverse Transcription Quantitative Real-Time PCR (RT-qPCR) ... For detection with the miRCURY LNA hsa-miR-222-3p (YP00204551 – targeted to the 21 nt isoform), 1.125 pmol of synthetic miRNA was polyadenylated and reverse transcribed with the miRCURY LNA RT Kit, following the manufacturer’s instructions (Qiagen).

SYBR Green Assay:

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: microRNA regulation of persistent stress-enhanced memory
Article Snippet: .. QPCR: A cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) and PCR reactions were performed in triplicate for each sample using the miRCURY LNA SYBR Green PCR Kit and the following locked nucleic acid (LNA) SYBR green primers from Qiagen. ..

Article Title: Down-regulation of long non-coding RNA MEG3 suppresses osteogenic differentiation of periodontal ligament stem cells (PDLSCs) through miR-27a-3p/IGF1 axis in periodontitis
Article Snippet: Thereafter, about 2 μg of total RNA was reversely transcripted to cDNA using SuperScriptTM III Reverse Transcriptase Kit (Invitrogen) for lncRNA/mRNA reverse transcription and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) for miRNA reverse transcription. .. Subsequently, qRT-PCR was then performed using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan) on LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland).

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Transcriptomic profiling of cell-free and vesicular microRNAs from matched arterial and venous sera
Article Snippet: Four µl RNA were used as starting material for reverse transcription (miRCURY LNA RT Kit, Qiagen, Venlo, The Netherlands). .. After dilution of cDNA (1:30), qPCR was performed using the miRCURY LNA SYBR Green PCR Kit.

Article Title: Bioinformatic analysis of long-lasting transcriptional and translational changes in the basolateral amygdala following acute stress
Article Snippet: QPCR validation For validation of smRNA-seq, a cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) following the manufacturer’s protocol and technical replicate samples that were sequenced. cDNA was diluted 1:80 for PCR. .. PCR reactions were performed in triplicate for each sample using the miRCURY LNA SYBR Green PCR Kit and the following locked nucleic acid (LNA) SYBR green primers from Qiagen (formerly Exiqon): mir-135b-3p, YP00204427; mir-199a-5p, YP00204494; mir-29a-5p, YP00204430.

Article Title: Modified Polyadenylation-Based RT-qPCR Increases Selectivity of Amplification of 3′-MicroRNA Isoforms
Article Snippet: The mixture was incubated for 1 h at 37°C and the reaction was stopped after incubation at 85°C for 5 min. Fifteen microliter of RNase and DNase free water was added to the reverse-transcribed polyadenylated total cellular RNA, and 1 μL of the resulting mix was used per qPCR reaction with the Power SYBR Green mastermix (Applied Biosystems). .. For detection with the miRCURY LNA hsa-miR-222-3p (YP00204551 – targeted to the 21 nt isoform), 1.125 pmol of synthetic miRNA was polyadenylated and reverse transcribed with the miRCURY LNA RT Kit, following the manufacturer’s instructions (Qiagen).

Article Title: Interplay between the Epigenetic Enzyme Lysine (K)-Specific Demethylase 2B and Epstein-Barr Virus Infection
Article Snippet: To analyze cellular miRNA, total RNA extracted by the AllPrep DNA/RNA minikit (Qiagen) was reverse transcribed using an miRCURY LNA miRNA PCR system (miRCURY LNA RT kit; catalog number 339340) according to the manufacturer’s protocol. .. The cDNA was then analyzed for the levels of miRNA-146-5p using a specific miRNA qPCR primer (catalog number 339306; Hsa-miR-146a-5p miRCURY LNA miRNA PCR assay) and a PCR kit (catalog number 339345; miRCURY LNA SYBR green PCR kits) on a Bio-Rad qPCR machine (CFX96 Touch real-time PCR).

Incubation:

Expressing:

Article Title: Reprogramming of Small Noncoding RNA Populations in Peripheral Blood Reveals Host Biomarkers for Latent and Active Mycobacterium tuberculosis Infection
Article Snippet: Paragraph title: Targeted quantification of let-7a-5p, miR-196b-5p, miR-589-5p, and SNORD104 expression. ... RNA yield was measured with a NanoDrop S1000 spectrophotometer (Thermo Scientific), and 50 ng was then reverse transcribed using a miRCURY LNA RT kit (Qiagen).

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Down-regulation of long non-coding RNA MEG3 suppresses osteogenic differentiation of periodontal ligament stem cells (PDLSCs) through miR-27a-3p/IGF1 axis in periodontitis
Article Snippet: Thereafter, about 2 μg of total RNA was reversely transcripted to cDNA using SuperScriptTM III Reverse Transcriptase Kit (Invitrogen) for lncRNA/mRNA reverse transcription and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) for miRNA reverse transcription. .. Normalization of mRNA and lncRNA expression values was performed against GAPDH and miRNA expression value was normalized against U6.

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

other:

Article Title: Effects on the Profile of Circulating miRNAs after Single Bouts of Resistance Training with and without Blood Flow Restriction—A Three-Arm, Randomized Crossover Trial
Article Snippet: Reverse Transcription and miRNA Profiling For reverse transcription, the miRCURY LNA RT Kit (Qiagen, Hilden, Germany) was used.

Polymerase Chain Reaction:

Article Title: Pancreatic β-cell tRNA hypomethylation and fragmentation link TRMT10A deficiency with diabetes
Article Snippet: .. After treatment, the RNA samples were polyadenylated and reverse transcribed using miRCURY LNA RT kit (Exiqon) and a poly(T) primer with a 3′ degenerate anchor and a 5′ universal tag. tRNA-derived fragments were quantified by real-time PCR using miRCURY LNATM SYBER Green PCR Kit (Exiqon), and specific custom-designed miRNA LNA™ PCR primers targeting the first 17 nucleotides of tRNAGln and hsa-let-7f-5p, used as housekeeping. .. Ten millon lymphoblasts were incubated for 15 min in 2 ml methionine- or glutamine-free RPMI-1640 supplemented with 10% dialyzed FBS, and then incubated for 10 min in the same medium containing 10 μCi/ml l -[35 S]-methionine or l -[3,4–3 H(N)]-glutamine at 37°C.

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Down-regulation of long non-coding RNA MEG3 suppresses osteogenic differentiation of periodontal ligament stem cells (PDLSCs) through miR-27a-3p/IGF1 axis in periodontitis
Article Snippet: Thereafter, about 2 μg of total RNA was reversely transcripted to cDNA using SuperScriptTM III Reverse Transcriptase Kit (Invitrogen) for lncRNA/mRNA reverse transcription and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) for miRNA reverse transcription. .. Subsequently, qRT-PCR was then performed using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan) on LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland).

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Bioinformatic analysis of long-lasting transcriptional and translational changes in the basolateral amygdala following acute stress
Article Snippet: .. QPCR validation For validation of smRNA-seq, a cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) following the manufacturer’s protocol and technical replicate samples that were sequenced. cDNA was diluted 1:80 for PCR. .. PCR reactions were performed in triplicate for each sample using the miRCURY LNA SYBR Green PCR Kit and the following locked nucleic acid (LNA) SYBR green primers from Qiagen (formerly Exiqon): mir-135b-3p, YP00204427; mir-199a-5p, YP00204494; mir-29a-5p, YP00204430.

Article Title: Interplay between the Epigenetic Enzyme Lysine (K)-Specific Demethylase 2B and Epstein-Barr Virus Infection
Article Snippet: .. To analyze cellular miRNA, total RNA extracted by the AllPrep DNA/RNA minikit (Qiagen) was reverse transcribed using an miRCURY LNA miRNA PCR system (miRCURY LNA RT kit; catalog number 339340) according to the manufacturer’s protocol. .. The cDNA was then analyzed for the levels of miRNA-146-5p using a specific miRNA qPCR primer (catalog number 339306; Hsa-miR-146a-5p miRCURY LNA miRNA PCR assay) and a PCR kit (catalog number 339345; miRCURY LNA SYBR green PCR kits) on a Bio-Rad qPCR machine (CFX96 Touch real-time PCR).

Recombinant:

Article Title: Pancreatic β-cell tRNA hypomethylation and fragmentation link TRMT10A deficiency with diabetes
Article Snippet: The recombinant His-tagged proteins were purified using HisLink protein purification resin (Promega). .. After treatment, the RNA samples were polyadenylated and reverse transcribed using miRCURY LNA RT kit (Exiqon) and a poly(T) primer with a 3′ degenerate anchor and a 5′ universal tag. tRNA-derived fragments were quantified by real-time PCR using miRCURY LNATM SYBER Green PCR Kit (Exiqon), and specific custom-designed miRNA LNA™ PCR primers targeting the first 17 nucleotides of tRNAGln and hsa-let-7f-5p, used as housekeeping.

RNA Sequencing Assay:

Article Title: Bioinformatic analysis of long-lasting transcriptional and translational changes in the basolateral amygdala following acute stress
Article Snippet: QPCR validation For validation of smRNA-seq, a cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) following the manufacturer’s protocol and technical replicate samples that were sequenced. cDNA was diluted 1:80 for PCR. .. For validation of RNA-seq, cDNA synthesis and qPCR were performed as previously described[ ] with 50ng of RNA using the following Taqman Assays (Thermo-Life Scientific): Chm, Mm00517015_m1; Dnajb4, Mm00508908_m1; Gabrb2, Mm00433467_m1; Rpl15, Mm01727317_g1; Slc8a1, Mm01232254_m1; Ube2d3, Mm00787086_s1.

Mutagenesis:

Article Title: Pancreatic β-cell tRNA hypomethylation and fragmentation link TRMT10A deficiency with diabetes
Article Snippet: Before reverse transcription, total RNA samples were subjected to 2′–3′ cyclic phosphate removal and 5′ hydroxyl group phosphorylation using rtStar™ tRF and tiRNA pretreatment Kit (ArrayStar), followed by m1 A, m3 C and m1 G demethylation using recombinant Escherichia coli AlkB demethylase, and its mutant AlkB-D135S ( ). .. After treatment, the RNA samples were polyadenylated and reverse transcribed using miRCURY LNA RT kit (Exiqon) and a poly(T) primer with a 3′ degenerate anchor and a 5′ universal tag. tRNA-derived fragments were quantified by real-time PCR using miRCURY LNATM SYBER Green PCR Kit (Exiqon), and specific custom-designed miRNA LNA™ PCR primers targeting the first 17 nucleotides of tRNAGln and hsa-let-7f-5p, used as housekeeping.

Isolation:

Article Title: Identification of stably expressed housekeeping miRNAs in endothelial cells and macrophages in an inflammatory setting
Article Snippet: Paragraph title: RNA isolation ... Complementary DNA (cDNA) was synthesized by means of the miRCURY LNA RT Kit (QIAGEN, Hilden, Germany) according to standard protocol.

Purification:

Protein Purification:

Sequencing:

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. PCR primers were synthesized by Sangon Biotech (Shanghai, China), and sequence information is exhibited in Table .

cDNA Library Assay:

Article Title: Bioinformatic analysis of long-lasting transcriptional and translational changes in the basolateral amygdala following acute stress
Article Snippet: .. QPCR validation For validation of smRNA-seq, a cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) following the manufacturer’s protocol and technical replicate samples that were sequenced. cDNA was diluted 1:80 for PCR. .. PCR reactions were performed in triplicate for each sample using the miRCURY LNA SYBR Green PCR Kit and the following locked nucleic acid (LNA) SYBR green primers from Qiagen (formerly Exiqon): mir-135b-3p, YP00204427; mir-199a-5p, YP00204494; mir-29a-5p, YP00204430.

Real-time Polymerase Chain Reaction:

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Long non-coding RNA HIF1A-AS2 facilitates adipose-derived stem cells (ASCs) osteogenic differentiation through miR-665/IL6 axis via PI3K/Akt signaling pathway
Article Snippet: .. qRT-PCR RNAs of ASCs were extracted using a TRIzol reagent (Invitrogen) and then reverse transcribed to cDNA using a SuperScript™ III Reverse Transcriptase Kit (Invitrogen) (for lncRNA and mRNA reverse transcription) and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) (for miRNA reverse transcription). qRT-PCR was conducted by a LightCycler 480 PCR System (Roche, Rotkreuz, Switzerland) using SYBR Green qPCR Master Mix (Takara, Tokyo, Japan). .. Relative expression of RNAs was calculated by a 2−ΔΔCt method.

Article Title: Bioinformatic analysis of long-lasting transcriptional and translational changes in the basolateral amygdala following acute stress
Article Snippet: .. QPCR validation For validation of smRNA-seq, a cDNA library was created from 50ng of total RNA using the mirCURY LNA RT Kit (Qiagen, Germantown, MD) following the manufacturer’s protocol and technical replicate samples that were sequenced. cDNA was diluted 1:80 for PCR. .. PCR reactions were performed in triplicate for each sample using the miRCURY LNA SYBR Green PCR Kit and the following locked nucleic acid (LNA) SYBR green primers from Qiagen (formerly Exiqon): mir-135b-3p, YP00204427; mir-199a-5p, YP00204494; mir-29a-5p, YP00204430.

Article Title: MicroRNA-deficient mouse embryonic stem cells acquire a functional interferon response
Article Snippet: RNA was first converted to cDNA using miRCURY LNA RT kit (Qiagen). cDNA was diluted 1/25 for RT-qPCR using miRCURY LNA SYBR Green kit and amplified using mmu-mmiR-673–5p specific primers (Qiagen) and U6 as a loading control. .. Quantitative PCR was carried out on a Roche LC480 light cycler and analysed using the second derivative method.

RNA Extraction:

Article Title: Reprogramming of Small Noncoding RNA Populations in Peripheral Blood Reveals Host Biomarkers for Latent and Active Mycobacterium tuberculosis Infection
Article Snippet: Total RNA was extracted from cells by the use of Qiazol reagent (Qiagen) and an miRNeasy RNA extraction kit (Qiagen), using on-column RNase-free DNase (Qiagen) treatment. .. RNA yield was measured with a NanoDrop S1000 spectrophotometer (Thermo Scientific), and 50 ng was then reverse transcribed using a miRCURY LNA RT kit (Qiagen).

RNA Expression:

Article Title: Down-regulation of long non-coding RNA MEG3 suppresses osteogenic differentiation of periodontal ligament stem cells (PDLSCs) through miR-27a-3p/IGF1 axis in periodontitis
Article Snippet: Thereafter, about 2 μg of total RNA was reversely transcripted to cDNA using SuperScriptTM III Reverse Transcriptase Kit (Invitrogen) for lncRNA/mRNA reverse transcription and miRCURY LNA RT Kit (Qiagen, Duesseldorf, Germany) for miRNA reverse transcription. .. Relative RNA expression was calculated with 2-ΔΔCt method.

Next-Generation Sequencing:

Article Title: Transcriptomic profiling of cell-free and vesicular microRNAs from matched arterial and venous sera
Article Snippet: RT-qPCR All miRNAs remaining differentially regulated after filtering the crude cell-free NGS data were tested in the same study population by RT-qPCR as described previously [ ]. .. Four µl RNA were used as starting material for reverse transcription (miRCURY LNA RT Kit, Qiagen, Venlo, The Netherlands).

Spectrophotometry:

Article Title: Reprogramming of Small Noncoding RNA Populations in Peripheral Blood Reveals Host Biomarkers for Latent and Active Mycobacterium tuberculosis Infection
Article Snippet: .. RNA yield was measured with a NanoDrop S1000 spectrophotometer (Thermo Scientific), and 50 ng was then reverse transcribed using a miRCURY LNA RT kit (Qiagen). .. Quantitative real-time PCR (RT-qPCR) was performed using a SensiFAST SYBR kit (Bioline).