mirna 3 0 genechip arrays  (Thermo Fisher)


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    Name:
    siRNA and miRNA Transfection Reagents
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    mirna
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    Thermo Fisher mirna 3 0 genechip arrays

    https://www.bioz.com/result/mirna 3 0 genechip arrays/product/Thermo Fisher
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mirna 3 0 genechip arrays - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: For generation of reporter plasmids, the identified human or porcine CAV2 target sites were obtained from hybridised oligonucleotides (Metabion AG) and were cloned in pTK-Gluc (NEB GmbH) and endotoxin-free reporter plasmids (pTKGhCAV2 and pTKGsCAV2) produced for transfection as described earlier . .. The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    Article Title: Regulation of Human RNase-L by the miR-29 Family Reveals a Novel Oncogenic Role in Chronic Myelogenous Leukemia
    Article Snippet: The RNase-L ZC5+ construct was then partially digested with Eco RI, and the 3′ UTR fragment was gel-purified and cloned into the Eco RI site downstream of firefly luciferase. .. Constructs were verified by sequencing (Biopolymer/Genomics Core Facility, University of Maryland, Baltimore). pGIPZ-encoded nonspecific and RNase-L short-hairpin RNAs (shRNAs) were purchased from Open Biosystems. miR-29 Pre-miR miRNA precursors and mirVana miRNA inhibitors were purchased from Ambion.

    Luciferase:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: Paragraph title: Transfection, Luciferase Reporter Assay, RNAi and Overexpression ... The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    Article Title: Regulation of Human RNase-L by the miR-29 Family Reveals a Novel Oncogenic Role in Chronic Myelogenous Leukemia
    Article Snippet: The Renilla luciferase plasmid was generously provided by Dr. Myriam Gorospe (National Institute on Aging, Baltimore). miR-29-site mutations were created using the QuikChange Site-Directed Mutagenesis Kit (Stratagene) using primers that deleted the 7 nucleotides predicted to bind the seed region (nt2–8) of the miR-29 family. .. Constructs were verified by sequencing (Biopolymer/Genomics Core Facility, University of Maryland, Baltimore). pGIPZ-encoded nonspecific and RNase-L short-hairpin RNAs (shRNAs) were purchased from Open Biosystems. miR-29 Pre-miR miRNA precursors and mirVana miRNA inhibitors were purchased from Ambion.

    Reporter Assay:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: Paragraph title: Transfection, Luciferase Reporter Assay, RNAi and Overexpression ... The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    Synthesized:

    Article Title: miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway
    Article Snippet: Total RNA was extracted using TRIzol reagent (Invitrogen, Paisley, UK) and then was treated with DNase I (Ambion, Paisley, UK) at 37 °C for 30 min. cDNAs were synthesized using a high-capacity cDNA reverse transcription Kit (Applied Biosystems, Foster City, CA, USA). .. For the qRT-PCR analysis of miR-216a, the TaqMan miRNA reverse transcription kit and TaqMan miRNA assay kit (Applied Biosystems) were used.

    Quantitative RT-PCR:

    Article Title: miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway
    Article Snippet: .. For the qRT-PCR analysis of miR-216a, the TaqMan miRNA reverse transcription kit and TaqMan miRNA assay kit (Applied Biosystems) were used. .. The relative expression of mRNA or miRNA was evaluated by the 2–ΔΔCt method and normalized to the expression of GAPDH or U6, respectively.

    Article Title: Circulating microRNAs in Pancreatic Juice as Candidate Biomarkers of Pancreatic Cancer
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis Taqman miRNA assays (Applied Biosystems, Foster City, CA) were used to perform expression profiling of the miRNAs of interest. .. RNA was pretreated with DNase (Invitrogen).

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: Paragraph title: MicroRNA amplification, detection and quantification by quantitative RT-PCR ... The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX).

    Article Title: Runx2 /DICER/miRNA Pathway in Regulating Osteogenesis †
    Article Snippet: .. For miRNA analysis, total RNA was extracted using the miRNeasy Mini Kit (Qiagen), cDNA synthesis was performed with 1μg of total RNA, using an NCode miRNA First-Strand cDNA Synthesis Kit (Invitrogen) according to the manufacturer’s protocol. qRT- PCR was performed on a Bio-Rad iQ5 thermal cycler using an NCode Express SYBR GreenER miRNA qRT-PCR Kit (Invitrogen). .. Expression levels of PCR product amounts were evaluated by the comparative cycle threshold method using U6 snRNA as a control ( ).

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. MicroRNA amplification, detection and quantification by quantitative RT-PCR Real time RT-PCR analysis of the microRNAs was performed using the Taqman MicroRNA Reverse Transcription Kit and the Taqman MicroRNA Assay (Applied Biosystems, Foster City, CA). .. For reverse transcription, 5.84 μl of a mix containing 3 μl of the RT primer solution (final concentration: 50 nM ), 0.15 μl dNTP (1 mM), 1 μl Multiscribe Reverse Transcriptase (3.33 U/μl), 1.50 μl of 10X Buffer and 0.19 μl RNase inhibitor (0.25 U/μl) were added to 9.16 μl of plasma RNA (final volume: 15 μl).

    Real-time Polymerase Chain Reaction:

    Article Title: Systemically Circulating Viral and Tumor-Derived MicroRNAs in KSHV-Associated Malignancies
    Article Snippet: .. RNA was isolated from exosome-enriched samples and the miRNA expression profile was determined by qPCR using Taqman microRNA assays from human pool A v2.1 (Life Technologies). ..

    Article Title: MicroRNA Controlled Adenovirus Mediates Anti-Cancer Efficacy without Affecting Endogenous MicroRNA Activity
    Article Snippet: Samples were run on a Step One Plus real-time PCR system (Applied Biosystems). .. The microRNA let7a was used as an internal standard using the conditions described above and using the TaqMan MicroRNA Assay (Applied Biosystems) specific for mature let7a RNA (Part number: 4427975 Assay ID: 000377).

    Article Title: Circulating microRNAs in Pancreatic Juice as Candidate Biomarkers of Pancreatic Cancer
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis Taqman miRNA assays (Applied Biosystems, Foster City, CA) were used to perform expression profiling of the miRNAs of interest. .. RNA was pretreated with DNase (Invitrogen).

    Article Title: The isomiR-140-3p-regulated mevalonic acid pathway as a potential target for prevention of triple negative breast cancer
    Article Snippet: Complementary DNA (cDNA) was prepared using an iScript cDNA synthesis kit (Bio-Rad) according to the manufacturer’s instructions. qPCR was performed in triplicate on each sample using an SYBR Green-based PCR assay as described previously [ ]. .. Mature miR-140-3p-1 and miR-140-3p-2 were quantified by using TaqMan-based miRNA assays from Thermo Scientific according to the manufacturer’s instructions.

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions. .. Real-time PCR reactions were run in triplicate using the ABI 7900HT Fast Real-Time PCR System and data were collected using the Sequence Detection System 2.4 (SDS) software (Applied Biosystems).

    Microarray:

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: Microarray data were analysed using a two-class paired significant analysis of microarray, as well as Pearson's correlations to determine differential gene expression before and after treatment using or correlations between miRNAs and target genes, respectively. .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions.

    Quantitation Assay:

    Article Title: MicroRNA Controlled Adenovirus Mediates Anti-Cancer Efficacy without Affecting Endogenous MicroRNA Activity
    Article Snippet: Paragraph title: MicroRNA Quantitation ... The microRNA let7a was used as an internal standard using the conditions described above and using the TaqMan MicroRNA Assay (Applied Biosystems) specific for mature let7a RNA (Part number: 4427975 Assay ID: 000377).

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions. .. Expression was calculated using the absolute quantitation standard curve method.

    Amplification:

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX). ..

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. MicroRNA amplification, detection and quantification by quantitative RT-PCR Real time RT-PCR analysis of the microRNAs was performed using the Taqman MicroRNA Reverse Transcription Kit and the Taqman MicroRNA Assay (Applied Biosystems, Foster City, CA). .. For reverse transcription, 5.84 μl of a mix containing 3 μl of the RT primer solution (final concentration: 50 nM ), 0.15 μl dNTP (1 mM), 1 μl Multiscribe Reverse Transcriptase (3.33 U/μl), 1.50 μl of 10X Buffer and 0.19 μl RNase inhibitor (0.25 U/μl) were added to 9.16 μl of plasma RNA (final volume: 15 μl).

    Expressing:

    Article Title: Systemically Circulating Viral and Tumor-Derived MicroRNAs in KSHV-Associated Malignancies
    Article Snippet: .. RNA was isolated from exosome-enriched samples and the miRNA expression profile was determined by qPCR using Taqman microRNA assays from human pool A v2.1 (Life Technologies). ..

    Article Title: miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway
    Article Snippet: For the qRT-PCR analysis of miR-216a, the TaqMan miRNA reverse transcription kit and TaqMan miRNA assay kit (Applied Biosystems) were used. .. The relative expression of mRNA or miRNA was evaluated by the 2–ΔΔCt method and normalized to the expression of GAPDH or U6, respectively.

    Article Title: Circulating microRNAs in Pancreatic Juice as Candidate Biomarkers of Pancreatic Cancer
    Article Snippet: .. Quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) Analysis Taqman miRNA assays (Applied Biosystems, Foster City, CA) were used to perform expression profiling of the miRNAs of interest. .. RNA was pretreated with DNase (Invitrogen).

    Article Title: Runx2 /DICER/miRNA Pathway in Regulating Osteogenesis †
    Article Snippet: For miRNA analysis, total RNA was extracted using the miRNeasy Mini Kit (Qiagen), cDNA synthesis was performed with 1μg of total RNA, using an NCode miRNA First-Strand cDNA Synthesis Kit (Invitrogen) according to the manufacturer’s protocol. qRT- PCR was performed on a Bio-Rad iQ5 thermal cycler using an NCode Express SYBR GreenER miRNA qRT-PCR Kit (Invitrogen). .. Expression levels of PCR product amounts were evaluated by the comparative cycle threshold method using U6 snRNA as a control ( ).

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: Microarray data were analysed using a two-class paired significant analysis of microarray, as well as Pearson's correlations to determine differential gene expression before and after treatment using or correlations between miRNAs and target genes, respectively. .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions.

    Genome Wide:

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: For genome-wide expression analysis in peripheral blood samples, we used the Human HT-12 v4 Expression Bead Chip (Illumina). .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions.

    Transformation Assay:

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: Background adjustment using variance-stabilizing transformation correction and quantile normalization was performed using the FlexArray package implemented in R (version 1.6.3). .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions.

    Over Expression:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: Paragraph title: Transfection, Luciferase Reporter Assay, RNAi and Overexpression ... The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    Transfection:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: Paragraph title: Transfection, Luciferase Reporter Assay, RNAi and Overexpression ... The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    TaqMan microRNA Assay:

    Article Title: miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway
    Article Snippet: .. For the qRT-PCR analysis of miR-216a, the TaqMan miRNA reverse transcription kit and TaqMan miRNA assay kit (Applied Biosystems) were used. .. The relative expression of mRNA or miRNA was evaluated by the 2–ΔΔCt method and normalized to the expression of GAPDH or U6, respectively.

    Article Title: MicroRNA Controlled Adenovirus Mediates Anti-Cancer Efficacy without Affecting Endogenous MicroRNA Activity
    Article Snippet: .. The microRNA let7a was used as an internal standard using the conditions described above and using the TaqMan MicroRNA Assay (Applied Biosystems) specific for mature let7a RNA (Part number: 4427975 Assay ID: 000377). .. Relative standard curves were produced by ten-fold serial dilutions of liver RNA extracted from a mouse receiving PBS.

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX). ..

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. MicroRNA amplification, detection and quantification by quantitative RT-PCR Real time RT-PCR analysis of the microRNAs was performed using the Taqman MicroRNA Reverse Transcription Kit and the Taqman MicroRNA Assay (Applied Biosystems, Foster City, CA). .. For reverse transcription, 5.84 μl of a mix containing 3 μl of the RT primer solution (final concentration: 50 nM ), 0.15 μl dNTP (1 mM), 1 μl Multiscribe Reverse Transcriptase (3.33 U/μl), 1.50 μl of 10X Buffer and 0.19 μl RNase inhibitor (0.25 U/μl) were added to 9.16 μl of plasma RNA (final volume: 15 μl).

    Polymerase Chain Reaction:

    Article Title: MicroRNA Controlled Adenovirus Mediates Anti-Cancer Efficacy without Affecting Endogenous MicroRNA Activity
    Article Snippet: Reaction conditions were 30 minutes 16°C, 30 minutes 42°C, 5 minutes 4°C. cDNA (1.33 µl of RT reaction) was added to 1 µl 20X Mir122 Taqman MicroRNA Assay, 10 µl TaqMan 2X Universal PCR master mix and made up to 20 µl using nuclease free water. .. The microRNA let7a was used as an internal standard using the conditions described above and using the TaqMan MicroRNA Assay (Applied Biosystems) specific for mature let7a RNA (Part number: 4427975 Assay ID: 000377).

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX). ..

    Article Title: The isomiR-140-3p-regulated mevalonic acid pathway as a potential target for prevention of triple negative breast cancer
    Article Snippet: Complementary DNA (cDNA) was prepared using an iScript cDNA synthesis kit (Bio-Rad) according to the manufacturer’s instructions. qPCR was performed in triplicate on each sample using an SYBR Green-based PCR assay as described previously [ ]. .. Mature miR-140-3p-1 and miR-140-3p-2 were quantified by using TaqMan-based miRNA assays from Thermo Scientific according to the manufacturer’s instructions.

    Article Title: Runx2 /DICER/miRNA Pathway in Regulating Osteogenesis †
    Article Snippet: The evaluation of PCR product amounts relative differences was carried out by the comparative cycle threshold method using GAPDH as a control. .. For miRNA analysis, total RNA was extracted using the miRNeasy Mini Kit (Qiagen), cDNA synthesis was performed with 1μg of total RNA, using an NCode miRNA First-Strand cDNA Synthesis Kit (Invitrogen) according to the manufacturer’s protocol. qRT- PCR was performed on a Bio-Rad iQ5 thermal cycler using an NCode Express SYBR GreenER miRNA qRT-PCR Kit (Invitrogen).

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: MicroRNA amplification, detection and quantification by quantitative RT-PCR Real time RT-PCR analysis of the microRNAs was performed using the Taqman MicroRNA Reverse Transcription Kit and the Taqman MicroRNA Assay (Applied Biosystems, Foster City, CA). .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX).

    Sequencing:

    Article Title: Regulation of Human RNase-L by the miR-29 Family Reveals a Novel Oncogenic Role in Chronic Myelogenous Leukemia
    Article Snippet: .. Constructs were verified by sequencing (Biopolymer/Genomics Core Facility, University of Maryland, Baltimore). pGIPZ-encoded nonspecific and RNase-L short-hairpin RNAs (shRNAs) were purchased from Open Biosystems. miR-29 Pre-miR miRNA precursors and mirVana miRNA inhibitors were purchased from Ambion. .. 293T, HeLa, and MDA-MB-231 cells were passaged in the Dulbecco's modified Eagle's medium (Cellgro) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Atlanta Biologicals), antibiotic/antimycotic (Invitrogen), and 2.5 μg/mL Plasmocin (InvivoGen).

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions. .. Real-time PCR reactions were run in triplicate using the ABI 7900HT Fast Real-Time PCR System and data were collected using the Sequence Detection System 2.4 (SDS) software (Applied Biosystems).

    Isolation:

    Article Title: Systemically Circulating Viral and Tumor-Derived MicroRNAs in KSHV-Associated Malignancies
    Article Snippet: .. RNA was isolated from exosome-enriched samples and the miRNA expression profile was determined by qPCR using Taqman microRNA assays from human pool A v2.1 (Life Technologies). ..

    Article Title: Mycotoxin ochratoxin A disrupts renal development via a miR-731/prolactin receptor axis in zebrafish a miR-731/prolactin receptor axis in zebrafish †Electronic supplementary information (ESI) available. See DOI: 10.1039/c5tx00360a
    Article Snippet: .. Total RNAs, including mRNA and microRNA, were isolated at the designated time by using the TRIzol-reagent (Invitrogen) according to the protocols of the manufacturer and Mraz et al . .. The cDNA was obtained from 2 μg of total RNA using SuperScript III (Invitrogen, Carlsbad, CA), and PCR was followed using designed primers, including PRLRa , serpina1 (serpin peptidase inhibitor, clade A, member1, ), c-myc ( , official symbol in zebrafish: myca) and EF (eukaryotic translation elongation factor 1 α, ) (Table S1 ).

    Negative Control:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies). .. The nonsense miRNA Pre-miR miRNA Precursor Negative Control #1 (Life technologies) as well as reporter plasmids harbouring the mutagenised target sites (pTKGhCAV2m and pTKGsCAV2m) were used as controls for specificity of interactions.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions. .. Real-time PCR reactions were run in triplicate using the ABI 7900HT Fast Real-Time PCR System and data were collected using the Sequence Detection System 2.4 (SDS) software (Applied Biosystems).

    Construct:

    Article Title: Regulation of Human RNase-L by the miR-29 Family Reveals a Novel Oncogenic Role in Chronic Myelogenous Leukemia
    Article Snippet: .. Constructs were verified by sequencing (Biopolymer/Genomics Core Facility, University of Maryland, Baltimore). pGIPZ-encoded nonspecific and RNase-L short-hairpin RNAs (shRNAs) were purchased from Open Biosystems. miR-29 Pre-miR miRNA precursors and mirVana miRNA inhibitors were purchased from Ambion. .. 293T, HeLa, and MDA-MB-231 cells were passaged in the Dulbecco's modified Eagle's medium (Cellgro) supplemented with 10% heat-inactivated fetal bovine serum (FBS; Atlanta Biologicals), antibiotic/antimycotic (Invitrogen), and 2.5 μg/mL Plasmocin (InvivoGen).

    Chromatin Immunoprecipitation:

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: For genome-wide expression analysis in peripheral blood samples, we used the Human HT-12 v4 Expression Bead Chip (Illumina). .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions.

    Plasmid Preparation:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: Nucleofection was performed using 5×105 –1×106 of either HeLa (Kit R) or HT-29 (Kit R) or IPEC-J2 (Kit L) using 1–2 µg reporter plasmid (pTK-Gluc derivatives, NEB GmbH), 100–200 ng normalisation plasmid (pTK-Cluc, NEB GmbH) and 100 pmol miRNA mimic according to the manufacturer’s instructions. .. The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    Article Title: Regulation of Human RNase-L by the miR-29 Family Reveals a Novel Oncogenic Role in Chronic Myelogenous Leukemia
    Article Snippet: The Renilla luciferase plasmid was generously provided by Dr. Myriam Gorospe (National Institute on Aging, Baltimore). miR-29-site mutations were created using the QuikChange Site-Directed Mutagenesis Kit (Stratagene) using primers that deleted the 7 nucleotides predicted to bind the seed region (nt2–8) of the miR-29 family. .. Constructs were verified by sequencing (Biopolymer/Genomics Core Facility, University of Maryland, Baltimore). pGIPZ-encoded nonspecific and RNase-L short-hairpin RNAs (shRNAs) were purchased from Open Biosystems. miR-29 Pre-miR miRNA precursors and mirVana miRNA inhibitors were purchased from Ambion.

    Software:

    Article Title: MicroRNAs 146a/b-5 and 425-3p and 24-3p are markers of antidepressant response and regulate MAPK/Wnt-system genes
    Article Snippet: All analyses were performed using MultiExperiment Viewer 4 (MeV4, TM4 software suite). .. For microRNA quantification, total RNA samples were reverse transcribed and quantified using TaqMan RT-PCR microRNA assays (Applied Biosystems) according to the manufacturer's instructions.

    SYBR Green Assay:

    Article Title: Runx2 /DICER/miRNA Pathway in Regulating Osteogenesis †
    Article Snippet: Bone metabolism genes mRNA analysis was performed by quantitative real-time reverse-transcriptase PCR (qRT-PCR) assay using iQ SYBR Green Supermix (Bio-Rad Laboratories, Hercules, CA, USA) on a Bio-Rad iQ5 thermal cycler (Bio-Rad Laboratories). .. For miRNA analysis, total RNA was extracted using the miRNeasy Mini Kit (Qiagen), cDNA synthesis was performed with 1μg of total RNA, using an NCode miRNA First-Strand cDNA Synthesis Kit (Invitrogen) according to the manufacturer’s protocol. qRT- PCR was performed on a Bio-Rad iQ5 thermal cycler using an NCode Express SYBR GreenER miRNA qRT-PCR Kit (Invitrogen).

    RNA Extraction:

    Article Title: miR-216a rescues dexamethasone suppression of osteogenesis, promotes osteoblast differentiation and enhances bone formation, by regulating c-Cbl-mediated PI3K/AKT pathway
    Article Snippet: Paragraph title: RNA extraction and qRT-PCR ... For the qRT-PCR analysis of miR-216a, the TaqMan miRNA reverse transcription kit and TaqMan miRNA assay kit (Applied Biosystems) were used.

    Article Title: The isomiR-140-3p-regulated mevalonic acid pathway as a potential target for prevention of triple negative breast cancer
    Article Snippet: Paragraph title: RNA extraction and quantitative (q)PCR ... Mature miR-140-3p-1 and miR-140-3p-2 were quantified by using TaqMan-based miRNA assays from Thermo Scientific according to the manufacturer’s instructions.

    Incubation:

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX). ..

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: MicroRNA amplification, detection and quantification by quantitative RT-PCR Real time RT-PCR analysis of the microRNAs was performed using the Taqman MicroRNA Reverse Transcription Kit and the Taqman MicroRNA Assay (Applied Biosystems, Foster City, CA). .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX).

    Produced:

    Article Title: Intestinal Salmonella typhimurium Infection Leads to miR-29a Induced Caveolin 2 Regulation
    Article Snippet: For generation of reporter plasmids, the identified human or porcine CAV2 target sites were obtained from hybridised oligonucleotides (Metabion AG) and were cloned in pTK-Gluc (NEB GmbH) and endotoxin-free reporter plasmids (pTKGhCAV2 and pTKGsCAV2) produced for transfection as described earlier . .. The reporter plasmids were co-transfected with Pre-miR miRNA Precursors miR-29a (Life Technologies).

    Article Title: MicroRNA Controlled Adenovirus Mediates Anti-Cancer Efficacy without Affecting Endogenous MicroRNA Activity
    Article Snippet: The microRNA let7a was used as an internal standard using the conditions described above and using the TaqMan MicroRNA Assay (Applied Biosystems) specific for mature let7a RNA (Part number: 4427975 Assay ID: 000377). .. Relative standard curves were produced by ten-fold serial dilutions of liver RNA extracted from a mouse receiving PBS.

    Concentration Assay:

    Article Title: MicroRNA Controlled Adenovirus Mediates Anti-Cancer Efficacy without Affecting Endogenous MicroRNA Activity
    Article Snippet: Briefly, Reverse transcription (RT) reactions (15 µl) were set up according to manufacturer's guidelines to using Multiscribe Reverse Transcriptase (50U/reaction), dNTPs (1 mM final concentration), 0.188 µl RNase inhibitor, 1.5 µl 10X RT buffer, 3 µl 5X mir122 TaqMan MicroRNA RT primer and 5 µl RNA sample (1 ng/µl). .. The microRNA let7a was used as an internal standard using the conditions described above and using the TaqMan MicroRNA Assay (Applied Biosystems) specific for mature let7a RNA (Part number: 4427975 Assay ID: 000377).

    Article Title: Consistent high concentration of the viral microRNA BART17 in plasma samples from nasopharyngeal carcinoma patients - evidence of non-exosomal transport
    Article Snippet: For reverse transcription, 5.84 μl of a mix containing 3 μl of the RT primer solution (final concentration: 50 nM ), 0.15 μl dNTP (1 mM), 1 μl Multiscribe Reverse Transcriptase (3.33 U/μl), 1.50 μl of 10X Buffer and 0.19 μl RNase inhibitor (0.25 U/μl) were added to 9.16 μl of plasma RNA (final volume: 15 μl). .. The mix was then incubated successively at 16°C for 30 minutes, 42°C for 30 min and 85°C for 5 min. After reverse transcription, PCR amplification was performed in 20 μl containing 4 μl of the final Reverse Transcription reaction mix mixed with 10 μl of FastStart Universal Probe Master mix (Roche diagnostics, Basel, Switzerland), 1 μl of the primer mix including - for a given microRNA - the universal reverse primer (0.7 μM), the specific primer (1.5 μM) and the hydrolysis probe (0.2 μM) (Taqman microRNA assay, Applied Biosystems) and 5 μl of nuclease free water (Ambion, Austin, TX).

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    Thermo Fisher genechip mirna 3 0
    General scheme of the miRNA profiling study design. In the discovery phase 10 patients were enrolled and analyzed through a <t>Genechip</t> miRNA 3.0 array. Patients were divided into two groups, with-HCC (subjects developing HCC after DAA treatment) and without-HCC (subjects not developing HCC after DAA treatment). Samples were analyzed at T0 and T1. Subsequently, miRNA candidates were assessed by qRT-PCR in 30 patients (validation cohort). MiRNA biomarkers were selected by estimating the time-averaged difference of a given miRNA between subjects with HCC vs. without HCC using a bootstrapped random-effect generalized least square regression model (RE-GLS).
    Genechip Mirna 3 0, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/genechip mirna 3 0/product/Thermo Fisher
    Average 99 stars, based on 3 article reviews
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    99
    Thermo Fisher whole genome microarray
    In Vitro Analysis of Off-Target Gene Regulation (A–F) Graphs showing hSOD1 mRNA level fold change as determined by qRT-PCR in HEK293T cells transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (A); isogenic Tet-inducible Flp-FRT HEK293 cells expressing either WT or G93A hSOD1 transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (B); iAstrocyte cells transduced with 250,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 10) (C); PCA plot of virally transduced isogenic Tet-inducible Flp-FRT HEK293 cell <t>microarray</t> data colored by treatment (scAAV9_hSOD1si, blue; scAAV9_hSOD1si_2, yellow; scAAV9_misSOD1si, pink; and scAAV9_H1emp, white) (D); volcano plots showing differentially expressed genes in scAAV9_hSOD1si transduced isogenic Tet-inducible Flp-FRT HEK293 cells (E) and iAstrocytes (F) compared to scAAV9_H1emp transduced cells (Probeset color key: green, p
    Whole Genome Microarray, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/whole genome microarray/product/Thermo Fisher
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    99
    Thermo Fisher microarray assay
    In Vitro Analysis of Off-Target Gene Regulation (A–F) Graphs showing hSOD1 mRNA level fold change as determined by qRT-PCR in HEK293T cells transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (A); isogenic Tet-inducible Flp-FRT HEK293 cells expressing either WT or G93A hSOD1 transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (B); iAstrocyte cells transduced with 250,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 10) (C); PCA plot of virally transduced isogenic Tet-inducible Flp-FRT HEK293 cell <t>microarray</t> data colored by treatment (scAAV9_hSOD1si, blue; scAAV9_hSOD1si_2, yellow; scAAV9_misSOD1si, pink; and scAAV9_H1emp, white) (D); volcano plots showing differentially expressed genes in scAAV9_hSOD1si transduced isogenic Tet-inducible Flp-FRT HEK293 cells (E) and iAstrocytes (F) compared to scAAV9_H1emp transduced cells (Probeset color key: green, p
    Microarray Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    General scheme of the miRNA profiling study design. In the discovery phase 10 patients were enrolled and analyzed through a Genechip miRNA 3.0 array. Patients were divided into two groups, with-HCC (subjects developing HCC after DAA treatment) and without-HCC (subjects not developing HCC after DAA treatment). Samples were analyzed at T0 and T1. Subsequently, miRNA candidates were assessed by qRT-PCR in 30 patients (validation cohort). MiRNA biomarkers were selected by estimating the time-averaged difference of a given miRNA between subjects with HCC vs. without HCC using a bootstrapped random-effect generalized least square regression model (RE-GLS).

    Journal: Cancers

    Article Title: Serum miRNA Are Promising Biomarkers for the Detection of Early Hepatocellular Carcinoma after Treatment with Direct-Acting Antivirals

    doi: 10.3390/cancers11111773

    Figure Lengend Snippet: General scheme of the miRNA profiling study design. In the discovery phase 10 patients were enrolled and analyzed through a Genechip miRNA 3.0 array. Patients were divided into two groups, with-HCC (subjects developing HCC after DAA treatment) and without-HCC (subjects not developing HCC after DAA treatment). Samples were analyzed at T0 and T1. Subsequently, miRNA candidates were assessed by qRT-PCR in 30 patients (validation cohort). MiRNA biomarkers were selected by estimating the time-averaged difference of a given miRNA between subjects with HCC vs. without HCC using a bootstrapped random-effect generalized least square regression model (RE-GLS).

    Article Snippet: Microarray Profiling 130 ng of purified small RNAs were labelled with the FlashTag™ Biotin HSR RNA Labeling Kit (Affymetrix® , Thermo Fischer Scientific) and hybridized on Genechip miRNA 3.0 (Thermo Fischer Scientific) containing 1734 human mature miRNAs.

    Techniques: Quantitative RT-PCR

    In Vitro Analysis of Off-Target Gene Regulation (A–F) Graphs showing hSOD1 mRNA level fold change as determined by qRT-PCR in HEK293T cells transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (A); isogenic Tet-inducible Flp-FRT HEK293 cells expressing either WT or G93A hSOD1 transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (B); iAstrocyte cells transduced with 250,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 10) (C); PCA plot of virally transduced isogenic Tet-inducible Flp-FRT HEK293 cell microarray data colored by treatment (scAAV9_hSOD1si, blue; scAAV9_hSOD1si_2, yellow; scAAV9_misSOD1si, pink; and scAAV9_H1emp, white) (D); volcano plots showing differentially expressed genes in scAAV9_hSOD1si transduced isogenic Tet-inducible Flp-FRT HEK293 cells (E) and iAstrocytes (F) compared to scAAV9_H1emp transduced cells (Probeset color key: green, p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Translating SOD1 Gene Silencing toward the Clinic: A Highly Efficacious, Off-Target-free, and Biomarker-Supported Strategy for fALS

    doi: 10.1016/j.omtn.2018.04.015

    Figure Lengend Snippet: In Vitro Analysis of Off-Target Gene Regulation (A–F) Graphs showing hSOD1 mRNA level fold change as determined by qRT-PCR in HEK293T cells transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (A); isogenic Tet-inducible Flp-FRT HEK293 cells expressing either WT or G93A hSOD1 transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (B); iAstrocyte cells transduced with 250,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 10) (C); PCA plot of virally transduced isogenic Tet-inducible Flp-FRT HEK293 cell microarray data colored by treatment (scAAV9_hSOD1si, blue; scAAV9_hSOD1si_2, yellow; scAAV9_misSOD1si, pink; and scAAV9_H1emp, white) (D); volcano plots showing differentially expressed genes in scAAV9_hSOD1si transduced isogenic Tet-inducible Flp-FRT HEK293 cells (E) and iAstrocytes (F) compared to scAAV9_H1emp transduced cells (Probeset color key: green, p

    Article Snippet: Microarray Analysis of Off-Target Effects Total RNA, extracted from Tet-inducible Flp-FRT HEK293 and iAstrocyte cells transduced with scAAV9 vectors expressing the off-target constructs (HEK293 cells, 50,000 vg/cell, collected 5 days post-transduction; iAstrocyte cells, 250,000 vg/cell, collected 5 days post-transduction), was analyzed by whole-genome microarray (HEK293 cells, 3′ IVT Express kit using GeneChip Human Genome U133 Plus 2.0 Arrays [Thermo Fisher Scientific]; iAstrocytes, GeneChip WT Plus reagent kit using Clariom S Human whole transcript arrays [Thermo Fisher Scientific]) to investigate gene expression changes elicited by treatment with the viral vectors.

    Techniques: In Vitro, Quantitative RT-PCR, Transduction, Expressing, Construct, Microarray

    In Vitro Analysis of Off-Target Gene Regulation (A–F) Graphs showing hSOD1 mRNA level fold change as determined by qRT-PCR in HEK293T cells transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (A); isogenic Tet-inducible Flp-FRT HEK293 cells expressing either WT or G93A hSOD1 transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (B); iAstrocyte cells transduced with 250,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 10) (C); PCA plot of virally transduced isogenic Tet-inducible Flp-FRT HEK293 cell microarray data colored by treatment (scAAV9_hSOD1si, blue; scAAV9_hSOD1si_2, yellow; scAAV9_misSOD1si, pink; and scAAV9_H1emp, white) (D); volcano plots showing differentially expressed genes in scAAV9_hSOD1si transduced isogenic Tet-inducible Flp-FRT HEK293 cells (E) and iAstrocytes (F) compared to scAAV9_H1emp transduced cells (Probeset color key: green, p

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Translating SOD1 Gene Silencing toward the Clinic: A Highly Efficacious, Off-Target-free, and Biomarker-Supported Strategy for fALS

    doi: 10.1016/j.omtn.2018.04.015

    Figure Lengend Snippet: In Vitro Analysis of Off-Target Gene Regulation (A–F) Graphs showing hSOD1 mRNA level fold change as determined by qRT-PCR in HEK293T cells transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (A); isogenic Tet-inducible Flp-FRT HEK293 cells expressing either WT or G93A hSOD1 transduced with 50,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 3) (B); iAstrocyte cells transduced with 250,000 vg/cell scAAV9 viral vectors expressing off-target constructs, harvested 120 hr post-transduction (n = 10) (C); PCA plot of virally transduced isogenic Tet-inducible Flp-FRT HEK293 cell microarray data colored by treatment (scAAV9_hSOD1si, blue; scAAV9_hSOD1si_2, yellow; scAAV9_misSOD1si, pink; and scAAV9_H1emp, white) (D); volcano plots showing differentially expressed genes in scAAV9_hSOD1si transduced isogenic Tet-inducible Flp-FRT HEK293 cells (E) and iAstrocytes (F) compared to scAAV9_H1emp transduced cells (Probeset color key: green, p

    Article Snippet: Total RNA, extracted from Tet-inducible Flp-FRT HEK293 and iAstrocyte cells transduced with scAAV9 vectors expressing the off-target constructs (HEK293 cells, 50,000 vg/cell, collected 5 days post-transduction; iAstrocyte cells, 250,000 vg/cell, collected 5 days post-transduction), was analyzed by whole-genome microarray (HEK293 cells, 3′ IVT Express kit using GeneChip Human Genome U133 Plus 2.0 Arrays [Thermo Fisher Scientific]; iAstrocytes, GeneChip WT Plus reagent kit using Clariom S Human whole transcript arrays [Thermo Fisher Scientific]) to investigate gene expression changes elicited by treatment with the viral vectors.

    Techniques: In Vitro, Quantitative RT-PCR, Transduction, Expressing, Construct, Microarray