ncode mirna first strand cdna synthesis kit  (Thermo Fisher)


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    Name:
    siRNA and miRNA Transfection Reagents
    Description:

    Catalog Number:
    mirna
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    Structured Review

    Thermo Fisher ncode mirna first strand cdna synthesis kit
    Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by <t>miRNA</t> QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first <t>cDNA</t> was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    https://www.bioz.com/result/ncode mirna first strand cdna synthesis kit/product/Thermo Fisher
    Average 90 stars, based on 73 article reviews
    Price from $9.99 to $1999.99
    ncode mirna first strand cdna synthesis kit - by Bioz Stars, 2020-02
    90/100 stars

    Images

    1) Product Images from "miR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells"

    Article Title: miR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S114276

    Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P
    Figure Legend Snippet: Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, Microscopy, Plasmid Preparation

    2) Product Images from "miR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells"

    Article Title: miR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells

    Journal: OncoTargets and therapy

    doi: 10.2147/OTT.S114276

    Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P
    Figure Legend Snippet: Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction, Microscopy, Plasmid Preparation

    Related Articles

    Amplification:

    Article Title: Tumor suppressive microRNA-137 negatively regulates Musashi-1 and colorectal cancer progression
    Article Snippet: Taqman microRNA analysis in patient tissue samples MicroRNA was isolated from paraffin-embedded tissue sections from rectal cancer patients using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Life Technologies, Stockholm, Sweden). .. The microRNA quality was assessed by studying amplification efficiencies of microRNAs after serial dilutions of cDNA and measuring CT values of microRNAs using real time PCR. miR-137 expression was determined using Taqman microRNA assay and normalized to control small RNA, RNU6b.

    Synthesized:

    Article Title: Role of microRNA-4458 in patients with non-small-cell lung cancer
    Article Snippet: Complementary (c)DNA was reverse transcribed using TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) on ice. .. Complementary (c)DNA was reverse transcribed using TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) on ice.

    Quantitative RT-PCR:

    Article Title: MicroRNA-328 Inhibits Renal Tubular Cell Epithelial-to-Mesenchymal Transition by Targeting the CD44 in Pressure-Induced Renal Fibrosis
    Article Snippet: .. For quantitative RT-PCR of miR-328, miRNAs in total RNA were reverse transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Grand Island, NY, USA), and then applied in TaqMan MicroRNA Assays with specific primers of rat miR-328 (cat. no. 4427975, miRBase ID: rno-miR-328a-3p) according to the manufacturer’s instructions (Life Technologies). .. Reactions were run on a real-time PCR system (ABI PRISMH 7700, Applied Biosystems, Austin, TX, USA).

    Article Title: miRNA Expression in Anaplastic Thyroid Carcinomas
    Article Snippet: .. Validation of miRNA expression data by quantitative real-time RT-PCR (qRT-PCR) qRT-PCR was performed using TaqMan MicroRNA Assay kits according to the manufacturer's protocol (Applied Biosystems). miRNA specific cDNA were generated from 10 ng of total RNA using the TaqMan microRNA RT kit and the gene-specific RT primers from the TaqMan microRNA Assays (Applied Biosystems) according to the manufacturer's protocol. ..

    Article Title: Expression profiles and circulation dynamics of rat mesenteric lymph microRNAs
    Article Snippet: .. RT-qPCR analysis RT-qPCR was performed using TaqMan MicroRNA assays (A and B; version 3.0; Thermo Fisher Scientific, Inc.) in a 7300 Real-Time PCR system (Thermo Fisher Scientific, Inc.) or a 7900HT Fast Real-Time PCR system (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. cel-miRNAs were used as exogenous controls for lymph and plasma samples to normalize miRNA expression levels, and U6 small nuclear RNA (Rnu6 ) was used as an endogenous internal control for tissue samples. .. Primers for miR-145 (assay ID 002278), miR-150 (assay ID 000473), cel-miR-39-3p (assay ID 000200), cel-miR-238-3p (assay ID 000248) and Rnu6 (assay ID 001973) were obtained from Thermo Fisher Scientific, Inc.

    Article Title: CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186
    Article Snippet: .. TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA) was used for miRNA reverse transcription and qRT-PCR was conducted using TaqMan Universal Master Mix II with TaqMan microRNA assays of miR-186-5p and U6 respectively. .. U6 and GAPDH were used as endogenous controls for miRNA and gene expression detection.

    Article Title: The function of miRNA-153 against isoflurane-induced neurotoxicity via Nrf2/ARE cytoprotection
    Article Snippet: .. Total RNA was reverse-transcribed into cDNA using a TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) at 37°C for 60 min and 85°C for 1 min. RT-qPCR was conducted using a Maxima SYBR Green PCR kit (Thermo Fisher Scientific Inc.). .. The following thermocycling conditions were used for qPCR: 95°C for 10 min; followed by 40 cycles at 95°C for 15 sec, 60°C for 30 sec and 72°C for 30 sec.

    Article Title: Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal
    Article Snippet: .. Reverse transcription and quantitative real time PCR (qRT-PCR) For miRNA detection, 10 ng of the RNA samples were reverse transcribed in technical duplicates using a TaqMan microRNA reverse transcription kit (Thermo Fisher Scientific) with Taqman microRNA primers specific for the miRNAs of interest, following manufacturer’s instructions (see Additional file : Table S1). .. Reverse transcription products were diluted to 50 μl and 5 μl of diluted sample used in single qPCR reactions, with a total volume of 20 μl. qRT-PCR was performed using Taqman microRNA assays (Thermo Fisher Scientific) and TaqMan universal PCR master mix II, no UNG (Thermo Fisher Scientific) on a ViiA7 Real-Time PCR system (Applied Biosystems/Thermo Fisher Scientific), using recommended PCR cycling conditions.

    Article Title: miR-205 enhances radiation sensitivity of prostate cancer cells by impairing DNA damage repair through PKCε and ZEB1 inhibition
    Article Snippet: .. Quantification of miR-205 and mRNA expression levels was assessed by qRT-PCR using the following TaqMan microRNA or gene expression assays (Thermo Fisher Scientific Inc): miR-205, 000,509; PKCε Hs00942886_m1 and ZEB1 Hs00232783_m1. .. Amplifications were run on the 7900HT Fast Real-Time PCR System.

    Article Title: Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas
    Article Snippet: Paragraph title: Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) for microRNA expression ... Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table ).

    Real-time Polymerase Chain Reaction:

    Article Title: MicroRNA-328 Inhibits Renal Tubular Cell Epithelial-to-Mesenchymal Transition by Targeting the CD44 in Pressure-Induced Renal Fibrosis
    Article Snippet: For quantitative RT-PCR of miR-328, miRNAs in total RNA were reverse transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Grand Island, NY, USA), and then applied in TaqMan MicroRNA Assays with specific primers of rat miR-328 (cat. no. 4427975, miRBase ID: rno-miR-328a-3p) according to the manufacturer’s instructions (Life Technologies). .. Reactions were run on a real-time PCR system (ABI PRISMH 7700, Applied Biosystems, Austin, TX, USA).

    Article Title: Tumor suppressive microRNA-137 negatively regulates Musashi-1 and colorectal cancer progression
    Article Snippet: Taqman microRNA analysis in patient tissue samples MicroRNA was isolated from paraffin-embedded tissue sections from rectal cancer patients using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Life Technologies, Stockholm, Sweden). .. The microRNA quality was assessed by studying amplification efficiencies of microRNAs after serial dilutions of cDNA and measuring CT values of microRNAs using real time PCR. miR-137 expression was determined using Taqman microRNA assay and normalized to control small RNA, RNU6b.

    Article Title: Expression profiles and circulation dynamics of rat mesenteric lymph microRNAs
    Article Snippet: .. RT-qPCR analysis RT-qPCR was performed using TaqMan MicroRNA assays (A and B; version 3.0; Thermo Fisher Scientific, Inc.) in a 7300 Real-Time PCR system (Thermo Fisher Scientific, Inc.) or a 7900HT Fast Real-Time PCR system (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. cel-miRNAs were used as exogenous controls for lymph and plasma samples to normalize miRNA expression levels, and U6 small nuclear RNA (Rnu6 ) was used as an endogenous internal control for tissue samples. .. Primers for miR-145 (assay ID 002278), miR-150 (assay ID 000473), cel-miR-39-3p (assay ID 000200), cel-miR-238-3p (assay ID 000248) and Rnu6 (assay ID 001973) were obtained from Thermo Fisher Scientific, Inc.

    Article Title: Role of microRNA-4458 in patients with non-small-cell lung cancer
    Article Snippet: Paragraph title: qPCR assay ... Complementary (c)DNA was reverse transcribed using TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) on ice.

    Article Title: miR-192 Mediates TGF-?/Smad3-Driven Renal Fibrosis
    Article Snippet: .. Total RNA (350 ng) was reverse-transcribed using TaqMan MicroRNA RT kit and Multiplex RT rodent primer pool (Applied Biosystems) according to the manufacturer's instructions. cDNAs were added to master mix (Applied Biosystems) and applied to miRNA TaqMan low-density array rodent panels 2.0 (Applied Biosystems) for the simultaneous quantification of 375 miRNAs according to the manufacturer's instructions. miRNA TaqMan assays were performed using 7900 HT Fast Real-Time PCR System (Applied Biosystems). .. Relative quantification was performed using ΔΔCt method, and the data were normalized with U6 and RNU48 (Applied Biosystems) as endogenous controls.

    Article Title: CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186
    Article Snippet: Paragraph title: Reverse transcription and quantitative real-time PCR (qRT-PCR) ... TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA) was used for miRNA reverse transcription and qRT-PCR was conducted using TaqMan Universal Master Mix II with TaqMan microRNA assays of miR-186-5p and U6 respectively.

    Article Title: The function of miRNA-153 against isoflurane-induced neurotoxicity via Nrf2/ARE cytoprotection
    Article Snippet: Total RNA was reverse-transcribed into cDNA using a TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) at 37°C for 60 min and 85°C for 1 min. RT-qPCR was conducted using a Maxima SYBR Green PCR kit (Thermo Fisher Scientific Inc.). .. The following thermocycling conditions were used for qPCR: 95°C for 10 min; followed by 40 cycles at 95°C for 15 sec, 60°C for 30 sec and 72°C for 30 sec.

    Article Title: Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal
    Article Snippet: .. Reverse transcription and quantitative real time PCR (qRT-PCR) For miRNA detection, 10 ng of the RNA samples were reverse transcribed in technical duplicates using a TaqMan microRNA reverse transcription kit (Thermo Fisher Scientific) with Taqman microRNA primers specific for the miRNAs of interest, following manufacturer’s instructions (see Additional file : Table S1). .. Reverse transcription products were diluted to 50 μl and 5 μl of diluted sample used in single qPCR reactions, with a total volume of 20 μl. qRT-PCR was performed using Taqman microRNA assays (Thermo Fisher Scientific) and TaqMan universal PCR master mix II, no UNG (Thermo Fisher Scientific) on a ViiA7 Real-Time PCR system (Applied Biosystems/Thermo Fisher Scientific), using recommended PCR cycling conditions.

    Article Title: miR-205 enhances radiation sensitivity of prostate cancer cells by impairing DNA damage repair through PKCε and ZEB1 inhibition
    Article Snippet: Quantification of miR-205 and mRNA expression levels was assessed by qRT-PCR using the following TaqMan microRNA or gene expression assays (Thermo Fisher Scientific Inc): miR-205, 000,509; PKCε Hs00942886_m1 and ZEB1 Hs00232783_m1. .. Amplifications were run on the 7900HT Fast Real-Time PCR System.

    Article Title: Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas
    Article Snippet: Paragraph title: Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) for microRNA expression ... Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table ).

    Microarray:

    Article Title: MicroRNA-328 Inhibits Renal Tubular Cell Epithelial-to-Mesenchymal Transition by Targeting the CD44 in Pressure-Induced Renal Fibrosis
    Article Snippet: Paragraph title: miRNA Microarray and Quantitative RT-PCR ... For quantitative RT-PCR of miR-328, miRNAs in total RNA were reverse transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Grand Island, NY, USA), and then applied in TaqMan MicroRNA Assays with specific primers of rat miR-328 (cat. no. 4427975, miRBase ID: rno-miR-328a-3p) according to the manufacturer’s instructions (Life Technologies).

    Article Title: Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas
    Article Snippet: Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) for microRNA expression Microarray expression of selected microRNAs was validated by RT-qPCR, both in the same cohort and in the validation cohort as described. .. Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table ).

    TLDA Assay:

    Article Title: miR-192 Mediates TGF-?/Smad3-Driven Renal Fibrosis
    Article Snippet: .. Total RNA (350 ng) was reverse-transcribed using TaqMan MicroRNA RT kit and Multiplex RT rodent primer pool (Applied Biosystems) according to the manufacturer's instructions. cDNAs were added to master mix (Applied Biosystems) and applied to miRNA TaqMan low-density array rodent panels 2.0 (Applied Biosystems) for the simultaneous quantification of 375 miRNAs according to the manufacturer's instructions. miRNA TaqMan assays were performed using 7900 HT Fast Real-Time PCR System (Applied Biosystems). .. Relative quantification was performed using ΔΔCt method, and the data were normalized with U6 and RNU48 (Applied Biosystems) as endogenous controls.

    Formalin-fixed Paraffin-Embedded:

    Article Title: Tumor suppressive microRNA-137 negatively regulates Musashi-1 and colorectal cancer progression
    Article Snippet: .. Taqman microRNA analysis in patient tissue samples MicroRNA was isolated from paraffin-embedded tissue sections from rectal cancer patients using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Life Technologies, Stockholm, Sweden). .. The microRNA quality was assessed by studying amplification efficiencies of microRNAs after serial dilutions of cDNA and measuring CT values of microRNAs using real time PCR. miR-137 expression was determined using Taqman microRNA assay and normalized to control small RNA, RNU6b.

    Expressing:

    Article Title: miRNA Expression in Anaplastic Thyroid Carcinomas
    Article Snippet: .. Validation of miRNA expression data by quantitative real-time RT-PCR (qRT-PCR) qRT-PCR was performed using TaqMan MicroRNA Assay kits according to the manufacturer's protocol (Applied Biosystems). miRNA specific cDNA were generated from 10 ng of total RNA using the TaqMan microRNA RT kit and the gene-specific RT primers from the TaqMan microRNA Assays (Applied Biosystems) according to the manufacturer's protocol. ..

    Article Title: Specific MicroRNA Pattern in Colon Tissue of Young Children with Eosinophilic Colitis
    Article Snippet: Total RNA was reverse-transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA, USA). .. MiRs were selected for validation according to the NGS results, and their expression was determined by RT-PCR using TaqMan Universal PCR Master Mix No AmpErase UNG (Life Technologies).

    Article Title: Tumor suppressive microRNA-137 negatively regulates Musashi-1 and colorectal cancer progression
    Article Snippet: Taqman microRNA analysis in patient tissue samples MicroRNA was isolated from paraffin-embedded tissue sections from rectal cancer patients using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Life Technologies, Stockholm, Sweden). .. The microRNA quality was assessed by studying amplification efficiencies of microRNAs after serial dilutions of cDNA and measuring CT values of microRNAs using real time PCR. miR-137 expression was determined using Taqman microRNA assay and normalized to control small RNA, RNU6b.

    Article Title: Expression profiles and circulation dynamics of rat mesenteric lymph microRNAs
    Article Snippet: .. RT-qPCR analysis RT-qPCR was performed using TaqMan MicroRNA assays (A and B; version 3.0; Thermo Fisher Scientific, Inc.) in a 7300 Real-Time PCR system (Thermo Fisher Scientific, Inc.) or a 7900HT Fast Real-Time PCR system (Thermo Fisher Scientific, Inc.) according to the manufacturer's protocol. cel-miRNAs were used as exogenous controls for lymph and plasma samples to normalize miRNA expression levels, and U6 small nuclear RNA (Rnu6 ) was used as an endogenous internal control for tissue samples. .. Primers for miR-145 (assay ID 002278), miR-150 (assay ID 000473), cel-miR-39-3p (assay ID 000200), cel-miR-238-3p (assay ID 000248) and Rnu6 (assay ID 001973) were obtained from Thermo Fisher Scientific, Inc.

    Article Title: miR-192 Mediates TGF-?/Smad3-Driven Renal Fibrosis
    Article Snippet: Paragraph title: miRNA Expression Analyses ... Total RNA (350 ng) was reverse-transcribed using TaqMan MicroRNA RT kit and Multiplex RT rodent primer pool (Applied Biosystems) according to the manufacturer's instructions. cDNAs were added to master mix (Applied Biosystems) and applied to miRNA TaqMan low-density array rodent panels 2.0 (Applied Biosystems) for the simultaneous quantification of 375 miRNAs according to the manufacturer's instructions. miRNA TaqMan assays were performed using 7900 HT Fast Real-Time PCR System (Applied Biosystems).

    Article Title: CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186
    Article Snippet: TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA) was used for miRNA reverse transcription and qRT-PCR was conducted using TaqMan Universal Master Mix II with TaqMan microRNA assays of miR-186-5p and U6 respectively. .. U6 and GAPDH were used as endogenous controls for miRNA and gene expression detection.

    Article Title: Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal
    Article Snippet: Reverse transcription and quantitative real time PCR (qRT-PCR) For miRNA detection, 10 ng of the RNA samples were reverse transcribed in technical duplicates using a TaqMan microRNA reverse transcription kit (Thermo Fisher Scientific) with Taqman microRNA primers specific for the miRNAs of interest, following manufacturer’s instructions (see Additional file : Table S1). .. For analysis of target gene mRNA expression 250 ng of RNA was reverse transcribed into cDNA using a Quantitect reverse transcription kit (Qiagen) in duplicate reactions.

    Article Title: miR-205 enhances radiation sensitivity of prostate cancer cells by impairing DNA damage repair through PKCε and ZEB1 inhibition
    Article Snippet: .. Quantification of miR-205 and mRNA expression levels was assessed by qRT-PCR using the following TaqMan microRNA or gene expression assays (Thermo Fisher Scientific Inc): miR-205, 000,509; PKCε Hs00942886_m1 and ZEB1 Hs00232783_m1. .. Amplifications were run on the 7900HT Fast Real-Time PCR System.

    Article Title: Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas
    Article Snippet: Paragraph title: Reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) for microRNA expression ... Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table ).

    Transfection:

    Article Title: MicroRNA response to hypoxic stress in soft tissue sarcoma cells: microRNA mediated regulation of HIF3?
    Article Snippet: .. Mimic transfections MiRIDIAN microRNA mimics (Thermo Scientific, Etten-Leur, Netherlands) of hsa-miR-210-3p and hsa-miR-485-5p and miRIDIAN microRNA Mimic Negative Control #1 (Thermo Scientific) were transfected in a final concentration of 50 nM using DharmaFECT1 transfection reagent (Thermo Scientific) 24 h after cell seeding. .. Transfection efficiency was optimized to > 95% using fluorescent mimics.

    TaqMan microRNA Assay:

    Article Title: miRNA Expression in Anaplastic Thyroid Carcinomas
    Article Snippet: .. Validation of miRNA expression data by quantitative real-time RT-PCR (qRT-PCR) qRT-PCR was performed using TaqMan MicroRNA Assay kits according to the manufacturer's protocol (Applied Biosystems). miRNA specific cDNA were generated from 10 ng of total RNA using the TaqMan microRNA RT kit and the gene-specific RT primers from the TaqMan microRNA Assays (Applied Biosystems) according to the manufacturer's protocol. ..

    Article Title: Tumor suppressive microRNA-137 negatively regulates Musashi-1 and colorectal cancer progression
    Article Snippet: Taqman microRNA analysis in patient tissue samples MicroRNA was isolated from paraffin-embedded tissue sections from rectal cancer patients using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Life Technologies, Stockholm, Sweden). .. The microRNA quality was assessed by studying amplification efficiencies of microRNAs after serial dilutions of cDNA and measuring CT values of microRNAs using real time PCR. miR-137 expression was determined using Taqman microRNA assay and normalized to control small RNA, RNU6b.

    Article Title: Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas
    Article Snippet: The RT-qPCR was done using a microRNA-specific TaqMan microRNA assay kit (Applied Biosystems, Foster City, CA) and Applied Biosystems 7900H Fast Real-Time system. .. Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table ).

    Generated:

    Article Title: miRNA Expression in Anaplastic Thyroid Carcinomas
    Article Snippet: .. Validation of miRNA expression data by quantitative real-time RT-PCR (qRT-PCR) qRT-PCR was performed using TaqMan MicroRNA Assay kits according to the manufacturer's protocol (Applied Biosystems). miRNA specific cDNA were generated from 10 ng of total RNA using the TaqMan microRNA RT kit and the gene-specific RT primers from the TaqMan microRNA Assays (Applied Biosystems) according to the manufacturer's protocol. ..

    other:

    Article Title: microRNA-605 promotes cell proliferation, migration and invasion in non-small cell lung cancer by directly targeting LATS2
    Article Snippet: TaqMan® microRNA assay (Applied Biosystems; Thermo Fisher Scientific, Inc.) was adopted to determine miR-650 expression, with U6 serving as an internal control.

    Polymerase Chain Reaction:

    Article Title: Specific MicroRNA Pattern in Colon Tissue of Young Children with Eosinophilic Colitis
    Article Snippet: Total RNA was reverse-transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA, USA). .. MiRs were selected for validation according to the NGS results, and their expression was determined by RT-PCR using TaqMan Universal PCR Master Mix No AmpErase UNG (Life Technologies).

    Article Title: The function of miRNA-153 against isoflurane-induced neurotoxicity via Nrf2/ARE cytoprotection
    Article Snippet: .. Total RNA was reverse-transcribed into cDNA using a TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) at 37°C for 60 min and 85°C for 1 min. RT-qPCR was conducted using a Maxima SYBR Green PCR kit (Thermo Fisher Scientific Inc.). .. The following thermocycling conditions were used for qPCR: 95°C for 10 min; followed by 40 cycles at 95°C for 15 sec, 60°C for 30 sec and 72°C for 30 sec.

    Article Title: Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal
    Article Snippet: Reverse transcription and quantitative real time PCR (qRT-PCR) For miRNA detection, 10 ng of the RNA samples were reverse transcribed in technical duplicates using a TaqMan microRNA reverse transcription kit (Thermo Fisher Scientific) with Taqman microRNA primers specific for the miRNAs of interest, following manufacturer’s instructions (see Additional file : Table S1). .. Reverse transcription products were diluted to 50 μl and 5 μl of diluted sample used in single qPCR reactions, with a total volume of 20 μl. qRT-PCR was performed using Taqman microRNA assays (Thermo Fisher Scientific) and TaqMan universal PCR master mix II, no UNG (Thermo Fisher Scientific) on a ViiA7 Real-Time PCR system (Applied Biosystems/Thermo Fisher Scientific), using recommended PCR cycling conditions.

    Multiplex Assay:

    Article Title: miR-192 Mediates TGF-?/Smad3-Driven Renal Fibrosis
    Article Snippet: .. Total RNA (350 ng) was reverse-transcribed using TaqMan MicroRNA RT kit and Multiplex RT rodent primer pool (Applied Biosystems) according to the manufacturer's instructions. cDNAs were added to master mix (Applied Biosystems) and applied to miRNA TaqMan low-density array rodent panels 2.0 (Applied Biosystems) for the simultaneous quantification of 375 miRNAs according to the manufacturer's instructions. miRNA TaqMan assays were performed using 7900 HT Fast Real-Time PCR System (Applied Biosystems). .. Relative quantification was performed using ΔΔCt method, and the data were normalized with U6 and RNU48 (Applied Biosystems) as endogenous controls.

    Isolation:

    Article Title: MicroRNA-328 Inhibits Renal Tubular Cell Epithelial-to-Mesenchymal Transition by Targeting the CD44 in Pressure-Induced Renal Fibrosis
    Article Snippet: The isolated total RNAs were then analyzed by Welgene Biotech (Taipei, Taiwan) with Agilent miRNA microarray (Sanger miRBase v.14). .. For quantitative RT-PCR of miR-328, miRNAs in total RNA were reverse transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Grand Island, NY, USA), and then applied in TaqMan MicroRNA Assays with specific primers of rat miR-328 (cat. no. 4427975, miRBase ID: rno-miR-328a-3p) according to the manufacturer’s instructions (Life Technologies).

    Article Title: Tumor suppressive microRNA-137 negatively regulates Musashi-1 and colorectal cancer progression
    Article Snippet: .. Taqman microRNA analysis in patient tissue samples MicroRNA was isolated from paraffin-embedded tissue sections from rectal cancer patients using RecoverAll™ Total Nucleic Acid Isolation Kit for FFPE (Life Technologies, Stockholm, Sweden). .. The microRNA quality was assessed by studying amplification efficiencies of microRNAs after serial dilutions of cDNA and measuring CT values of microRNAs using real time PCR. miR-137 expression was determined using Taqman microRNA assay and normalized to control small RNA, RNU6b.

    Size-exclusion Chromatography:

    Article Title: The function of miRNA-153 against isoflurane-induced neurotoxicity via Nrf2/ARE cytoprotection
    Article Snippet: Total RNA was reverse-transcribed into cDNA using a TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) at 37°C for 60 min and 85°C for 1 min. RT-qPCR was conducted using a Maxima SYBR Green PCR kit (Thermo Fisher Scientific Inc.). .. The following thermocycling conditions were used for qPCR: 95°C for 10 min; followed by 40 cycles at 95°C for 15 sec, 60°C for 30 sec and 72°C for 30 sec.

    Labeling:

    Article Title: Unique microRNA-profiles in EGFR-mutated lung adenocarcinomas
    Article Snippet: Reverse transcription of 10 ng of RNA was performed by employing the Taqman microRNA reverse transcription kit and microRNA-specific primers (Applied Biosystems) (Supporting Information Table ). .. The cDNA from the reverse transcription reaction was added to the TaqMan Universal Master Mix II no UNG and the labeled microRNA specific TaqMan probes (Applied Biosystems).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Specific MicroRNA Pattern in Colon Tissue of Young Children with Eosinophilic Colitis
    Article Snippet: Paragraph title: 4.4. RT-PCR ... Total RNA was reverse-transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA, USA).

    Article Title: CRNDE affects the malignant biological characteristics of human glioma stem cells by negatively regulating miR-186
    Article Snippet: One-Step SYBR PrimeScript RT-PCR Kit (Perfect Real Time) (Takara Bio, Inc, Japan) was used for qRT-PCR. .. TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems, Foster City, CA, USA) was used for miRNA reverse transcription and qRT-PCR was conducted using TaqMan Universal Master Mix II with TaqMan microRNA assays of miR-186-5p and U6 respectively.

    Software:

    Article Title: Role of microRNA-4458 in patients with non-small-cell lung cancer
    Article Snippet: Complementary (c)DNA was reverse transcribed using TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) on ice. .. Primer Design software version 5 (PREMIER Biosoft, Palo Alto, CA, USA) and DNAMAN gene analysis software demo version 5.2.9 (Lynnon Biosoft, San Ramon, CA, USA) were used in order to detect the primer location and to screen the appropriate primers.

    Article Title: miR-205 enhances radiation sensitivity of prostate cancer cells by impairing DNA damage repair through PKCε and ZEB1 inhibition
    Article Snippet: Quantification of miR-205 and mRNA expression levels was assessed by qRT-PCR using the following TaqMan microRNA or gene expression assays (Thermo Fisher Scientific Inc): miR-205, 000,509; PKCε Hs00942886_m1 and ZEB1 Hs00232783_m1. .. Data were analyzed by SDS 2.2.2 software (Thermo Fisher Scientific Inc) and reported as relative quantity with respect to a calibrator sample using the 2-∆∆Ct method.

    SYBR Green Assay:

    Article Title: The function of miRNA-153 against isoflurane-induced neurotoxicity via Nrf2/ARE cytoprotection
    Article Snippet: .. Total RNA was reverse-transcribed into cDNA using a TaqMan MicroRNA Reverse Transcription kit (Applied Biosystems; Thermo Fisher Scientific, Inc.) at 37°C for 60 min and 85°C for 1 min. RT-qPCR was conducted using a Maxima SYBR Green PCR kit (Thermo Fisher Scientific Inc.). .. The following thermocycling conditions were used for qPCR: 95°C for 10 min; followed by 40 cycles at 95°C for 15 sec, 60°C for 30 sec and 72°C for 30 sec.

    Negative Control:

    Article Title: MicroRNA response to hypoxic stress in soft tissue sarcoma cells: microRNA mediated regulation of HIF3?
    Article Snippet: .. Mimic transfections MiRIDIAN microRNA mimics (Thermo Scientific, Etten-Leur, Netherlands) of hsa-miR-210-3p and hsa-miR-485-5p and miRIDIAN microRNA Mimic Negative Control #1 (Thermo Scientific) were transfected in a final concentration of 50 nM using DharmaFECT1 transfection reagent (Thermo Scientific) 24 h after cell seeding. .. Transfection efficiency was optimized to > 95% using fluorescent mimics.

    Next-Generation Sequencing:

    Article Title: Specific MicroRNA Pattern in Colon Tissue of Young Children with Eosinophilic Colitis
    Article Snippet: Total RNA was reverse-transcribed using TaqMan MicroRNA Reverse Transcription Kit (Life Technologies, Carlsbad, CA, USA). .. MiRs were selected for validation according to the NGS results, and their expression was determined by RT-PCR using TaqMan Universal PCR Master Mix No AmpErase UNG (Life Technologies).

    Concentration Assay:

    Article Title: MicroRNA response to hypoxic stress in soft tissue sarcoma cells: microRNA mediated regulation of HIF3?
    Article Snippet: .. Mimic transfections MiRIDIAN microRNA mimics (Thermo Scientific, Etten-Leur, Netherlands) of hsa-miR-210-3p and hsa-miR-485-5p and miRIDIAN microRNA Mimic Negative Control #1 (Thermo Scientific) were transfected in a final concentration of 50 nM using DharmaFECT1 transfection reagent (Thermo Scientific) 24 h after cell seeding. .. Transfection efficiency was optimized to > 95% using fluorescent mimics.

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    Thermo Fisher ncode mirna first strand cdna synthesis kit
    Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by <t>miRNA</t> QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first <t>cDNA</t> was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P
    Ncode Mirna First Strand Cdna Synthesis Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    Journal: OncoTargets and therapy

    Article Title: miR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells

    doi: 10.2147/OTT.S114276

    Figure Lengend Snippet: Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    Article Snippet: In brief, total RNA was isolated using TRIzol reagent (Thermo Fisher Scientific), reverse transcribed with NCode™ miRNA First-Strand cDNA synthesis Kit (Thermo-Fisher Scientific).

    Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Microscopy, Plasmid Preparation

    Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    Journal: OncoTargets and therapy

    Article Title: miR-99a-5p acts as tumor suppressor via targeting to mTOR and enhances RAD001-induced apoptosis in human urinary bladder urothelial carcinoma cells

    doi: 10.2147/OTT.S114276

    Figure Lengend Snippet: Elevation of miR-99a-5p expression levels in 5637 and T24 cells transfected with pSM-99a. Notes: ( A ) miR-99a-5p was downregulated in 5637 and T24 cells. Cells at 70% confluence were collected, and the expression of miR-99a-5p was detected by miRNA QPCR. ( B and C ) Transfection of pSM-99a elevated the expression level of miR-99a-5p in 5637 and T24 cells. Transfected cells were selected with 200 μg/mL G418 for 24 h, while co-expressed EGFP was detectable under fluorescent microscopy. Total RNA was extracted and reverse transcribed, and the resulting first cDNA was applied to the subsequent miRNA QPCR. miR-99a-5p expression was normalized by U6 RNA. Data represent three independent experiments and are expressed as fold ± SD relative to the SV-Huc1 or vector only control (pSM). * P

    Article Snippet: In brief, total RNA was isolated using TRIzol reagent (Thermo Fisher Scientific), reverse transcribed with NCode™ miRNA First-Strand cDNA synthesis Kit (Thermo-Fisher Scientific).

    Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Microscopy, Plasmid Preparation