wt expression kit  (Thermo Fisher)


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    Name:
    WT Expression Kit
    Description:
    The WT Expression Kit is an RNA amplification kit that is designed to generate amplified sense strand cDNA ready for fragmentation and labeling using the Affymetrix GeneChip WT Terminal Labeling Kit Features of the WT Expression Kit • Optimized for use with Affymetrix GeneChip Human Mouse and Rat 1 0 ST Arrays• Allows RNA samples as small as 50 ng to be analyzed on Affymetrix GeneChip Arrays• Supports a streamlined workflow that does not require a separate rRNA depletion step• Is ideal for high sensitivity expression profilingEliminates separate rRNA depletion step preserves transcriptome coverageThe first generation Affymetrix amplification method requires depletion of ribosomal RNA rRNA from RNA samples for optimal exon level analysis Conversely the Ambion WT Expression Kit uses a novel reverse transcription RT priming method that eliminates the need for a separate rRNA depletion step The kit includes RT primers designed using a proprietary oligonucleotide matching algorithm that eliminates primer sequences with homology to known ribosomal RNAs The result is complete and unbiased coverage of the transcriptome with rRNA amplification levels significantly lower than those of other methods Consistent results from less input RNAWith the Ambion WT Expression Kit samples as small as 50 ng of total RNA can be analyzed on Affymetrix GeneChip Human Mouse and Rat Exon and Gene 1 0 ST Arrays Previous methods required 1 µg of total RNA often difficult or impossible to obtain from limited sources such as stem cells or small tissue samples The modest requirement for input RNA permits the analysis of rare samples and provides for more efficient and cost effective experimentation for most sample types To demonstrate the performance of the WT Expression Kit RNA from three sample types HeLa cells and Microarray Quality Control MAQC A and B samples was prepared in triplicate using either the Affymetrix GeneChip WT cDNA Synthesis and Amplification Kit or the WT Expression Kit and analyzed on Human Exon 1 0 ST Arrays Total RNA 1 µg prepared by the Affymetrix protocol underwent an rRNA depletion step while just 50 ng total RNA 20 fold less was prepared for microarray analysis using the WT Expression Kit Both sets of samples were handled according to the manufacturers recommendations Direct correlation of log2 ratios for MAQC samples A and B was high r 0 94 at both the exon and transcript levels Correlation of log2 ratios with gene expression data obtained using TaqMan Gene Expression assays for real time PCR was also high for both kits see Product Bulletin for data Identify more differential expression with less RNAIn addition to its lower input RNA requirement the WT Expression Kit provides a significant increase in sensitivity A greater number of probe sets detected above background was obtained at the exon level with the WT Expression Kit as a result of an increased signal to noise ratio A comparison of differential expression analysis results from the two methods indicated that while differentially expressed genes and exons of MAQC A and MAQC B samples were similar between the two kits 78 and 89 respectively significantly more differential expression was observed on arrays hybridized with samples prepared using the WT Expression Kit 499 more genes and 6 850 more exons were found to be differentially expressed in samples prepared using the Ambion kit compared to samples prepared with the Affymetrix kit Note For manual use only Not for robotics
    Catalog Number:
    4411973
    Price:
    None
    Applications:
    RNAi, Epigenetics & Non-Coding RNA Research|miRNA & Non-Coding RNA Analysis
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher wt expression kit
    The WT Expression Kit is an RNA amplification kit that is designed to generate amplified sense strand cDNA ready for fragmentation and labeling using the Affymetrix GeneChip WT Terminal Labeling Kit Features of the WT Expression Kit • Optimized for use with Affymetrix GeneChip Human Mouse and Rat 1 0 ST Arrays• Allows RNA samples as small as 50 ng to be analyzed on Affymetrix GeneChip Arrays• Supports a streamlined workflow that does not require a separate rRNA depletion step• Is ideal for high sensitivity expression profilingEliminates separate rRNA depletion step preserves transcriptome coverageThe first generation Affymetrix amplification method requires depletion of ribosomal RNA rRNA from RNA samples for optimal exon level analysis Conversely the Ambion WT Expression Kit uses a novel reverse transcription RT priming method that eliminates the need for a separate rRNA depletion step The kit includes RT primers designed using a proprietary oligonucleotide matching algorithm that eliminates primer sequences with homology to known ribosomal RNAs The result is complete and unbiased coverage of the transcriptome with rRNA amplification levels significantly lower than those of other methods Consistent results from less input RNAWith the Ambion WT Expression Kit samples as small as 50 ng of total RNA can be analyzed on Affymetrix GeneChip Human Mouse and Rat Exon and Gene 1 0 ST Arrays Previous methods required 1 µg of total RNA often difficult or impossible to obtain from limited sources such as stem cells or small tissue samples The modest requirement for input RNA permits the analysis of rare samples and provides for more efficient and cost effective experimentation for most sample types To demonstrate the performance of the WT Expression Kit RNA from three sample types HeLa cells and Microarray Quality Control MAQC A and B samples was prepared in triplicate using either the Affymetrix GeneChip WT cDNA Synthesis and Amplification Kit or the WT Expression Kit and analyzed on Human Exon 1 0 ST Arrays Total RNA 1 µg prepared by the Affymetrix protocol underwent an rRNA depletion step while just 50 ng total RNA 20 fold less was prepared for microarray analysis using the WT Expression Kit Both sets of samples were handled according to the manufacturers recommendations Direct correlation of log2 ratios for MAQC samples A and B was high r 0 94 at both the exon and transcript levels Correlation of log2 ratios with gene expression data obtained using TaqMan Gene Expression assays for real time PCR was also high for both kits see Product Bulletin for data Identify more differential expression with less RNAIn addition to its lower input RNA requirement the WT Expression Kit provides a significant increase in sensitivity A greater number of probe sets detected above background was obtained at the exon level with the WT Expression Kit as a result of an increased signal to noise ratio A comparison of differential expression analysis results from the two methods indicated that while differentially expressed genes and exons of MAQC A and MAQC B samples were similar between the two kits 78 and 89 respectively significantly more differential expression was observed on arrays hybridized with samples prepared using the WT Expression Kit 499 more genes and 6 850 more exons were found to be differentially expressed in samples prepared using the Ambion kit compared to samples prepared with the Affymetrix kit Note For manual use only Not for robotics
    https://www.bioz.com/result/wt expression kit/product/Thermo Fisher
    Average 99 stars, based on 1017 article reviews
    Price from $9.99 to $1999.99
    wt expression kit - by Bioz Stars, 2021-01
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Context-dependent anticodon recognition by class I lysyl-tRNA synthetases
    Article Snippet: .. Expand High Fidelity PCR system was from Roche, pBAD-TOPO cloning and expression kit was from Invitrogen, Ni-NTA matrix was from Qiagen (Chatsworth, CA), and L(+) arabinose was from Sigma. .. Mature Escherichia coli tRNALys was purchased from Sigma.

    Transfection:

    Article Title: KIT with D816 mutations cooperates with CBFB-MYH11 for leukemogenesis in mice
    Article Snippet: .. Phenix packaging cells were transfected with vectors carrying MSCV- KIT D816V, KIT D816Y, WT KIT , or green fluorescent protein (GFP) alone using a calcium transfection kit from Invitrogen. .. Two days after transfection, the supernatants were harvested and incubated with either the ecotropic packaging cell line GP+E-86 that was derived from mouse NIH3T3 cells, or 293T cells for 2 days.

    Amplification:

    Article Title: Nanostructural and Transcriptomic Analyses of Human Saliva Derived Exosomes
    Article Snippet: .. Briefly, single-stranded cDNA was generated from the amplified cRNA with the WT cDNA Synthesis Kit (Affymetrix, Santa Clara, CA) and then fragmented and labeled with the WT Terminal Labeling Kit (Affymetrix). .. Samples were hybridized with GeneChip HGU-133 plus 2 Arrays (Affymetrix) and scanned at the UCLA Microarray Core Facility.

    Article Title: Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats
    Article Snippet: .. Affymetrix GeneChip Rat Gene 1.0 ST Array System was used for the microarray analysis following the standard protocol: 100 ng RNA was amplified with Ambion WT Expression Kit (Applied Biosystems), 5.5 µg single-stranded cDNA was labeled and fragmented with GeneChip WT Terminal Labeling and Hybridization (Affymetrix) and hybridized on the chip according to the manufacturer procedure. .. Gene expression determined by real time PCR Total RNA was extracted from liver using Trizol reagent (Invitrogen), and cDNA was prepared and analyzed by real-time PCR testing using QuantiTect SYBR Green reagents (Qiagen, Inc.) on an Opticon continuous fluorescence detector (MJ Research).

    Isolation:

    Article Title: Spaceflight Activates Autophagy Programs and the Proteasome in Mouse Liver
    Article Snippet: .. Briefly, RNA was isolated using an RNeasy kit (Qiagen, Germantown, MD, USA) and the Ambion WT expression kit (Thermo Fisher Scientific, Waltham, MA, USA) was employed to prepare mRNA for whole transcriptome microarray analysis using an Affymetrix GeneChip 1.0 ST array (Thermo Fisher Scientific). .. Arrays were scanned using a GeneChip Scanner 3000 7G (Thermo Fisher Scientific) and Command Console Software v. 3.2.3 (Thermo Fisher Scientific) to produce.

    Labeling:

    Article Title: Nanostructural and Transcriptomic Analyses of Human Saliva Derived Exosomes
    Article Snippet: .. Briefly, single-stranded cDNA was generated from the amplified cRNA with the WT cDNA Synthesis Kit (Affymetrix, Santa Clara, CA) and then fragmented and labeled with the WT Terminal Labeling Kit (Affymetrix). .. Samples were hybridized with GeneChip HGU-133 plus 2 Arrays (Affymetrix) and scanned at the UCLA Microarray Core Facility.

    Article Title: Interhemispheric gene expression differences in the cerebral cortex of humans and macaque monkeys
    Article Snippet: .. Hybridization targets were prepared with an Ambion WT Expression Kit (Applied Biosystems/Ambion, Austin, TX) and Affymetrix GeneChip WT Terminal Labeling Kit from total RNA, hybridized to individual GeneChip® Human Gene 2.0 ST arrays in Affymetrix GeneChip® hybridization oven 645. .. Microarrays were washed in Affymetrix GeneChip® Fluidics Station 450 and scanned with Affymetrix GeneChip® Scanner 7G according to standard Affymetrix GeneChip® hybridization, wash, and stain protocols (Affymetrix, Santa Clara, CA).

    Article Title: Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats
    Article Snippet: .. Affymetrix GeneChip Rat Gene 1.0 ST Array System was used for the microarray analysis following the standard protocol: 100 ng RNA was amplified with Ambion WT Expression Kit (Applied Biosystems), 5.5 µg single-stranded cDNA was labeled and fragmented with GeneChip WT Terminal Labeling and Hybridization (Affymetrix) and hybridized on the chip according to the manufacturer procedure. .. Gene expression determined by real time PCR Total RNA was extracted from liver using Trizol reagent (Invitrogen), and cDNA was prepared and analyzed by real-time PCR testing using QuantiTect SYBR Green reagents (Qiagen, Inc.) on an Opticon continuous fluorescence detector (MJ Research).

    Microarray:

    Article Title: Spaceflight Activates Autophagy Programs and the Proteasome in Mouse Liver
    Article Snippet: .. Briefly, RNA was isolated using an RNeasy kit (Qiagen, Germantown, MD, USA) and the Ambion WT expression kit (Thermo Fisher Scientific, Waltham, MA, USA) was employed to prepare mRNA for whole transcriptome microarray analysis using an Affymetrix GeneChip 1.0 ST array (Thermo Fisher Scientific). .. Arrays were scanned using a GeneChip Scanner 3000 7G (Thermo Fisher Scientific) and Command Console Software v. 3.2.3 (Thermo Fisher Scientific) to produce.

    Article Title: Spheres Derived from Lung Adenocarcinoma Pleural Effusions: Molecular Characterization and Tumor Engraftment
    Article Snippet: .. Microarray Analysis Single strand biotinylated cDNA was generated as follows: 100 ng of total RNA were subjected to two cycles of cDNA synthesis with the Ambion WT expression Kit (Applera). ..

    Article Title: Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats
    Article Snippet: .. Affymetrix GeneChip Rat Gene 1.0 ST Array System was used for the microarray analysis following the standard protocol: 100 ng RNA was amplified with Ambion WT Expression Kit (Applied Biosystems), 5.5 µg single-stranded cDNA was labeled and fragmented with GeneChip WT Terminal Labeling and Hybridization (Affymetrix) and hybridized on the chip according to the manufacturer procedure. .. Gene expression determined by real time PCR Total RNA was extracted from liver using Trizol reagent (Invitrogen), and cDNA was prepared and analyzed by real-time PCR testing using QuantiTect SYBR Green reagents (Qiagen, Inc.) on an Opticon continuous fluorescence detector (MJ Research).

    Generated:

    Article Title: Nanostructural and Transcriptomic Analyses of Human Saliva Derived Exosomes
    Article Snippet: .. Briefly, single-stranded cDNA was generated from the amplified cRNA with the WT cDNA Synthesis Kit (Affymetrix, Santa Clara, CA) and then fragmented and labeled with the WT Terminal Labeling Kit (Affymetrix). .. Samples were hybridized with GeneChip HGU-133 plus 2 Arrays (Affymetrix) and scanned at the UCLA Microarray Core Facility.

    Article Title: Spheres Derived from Lung Adenocarcinoma Pleural Effusions: Molecular Characterization and Tumor Engraftment
    Article Snippet: .. Microarray Analysis Single strand biotinylated cDNA was generated as follows: 100 ng of total RNA were subjected to two cycles of cDNA synthesis with the Ambion WT expression Kit (Applera). ..

    Expressing:

    Article Title: Macrophage colony-stimulating factor (CSF1) controls monocyte production and maturation and the steady-state size of the liver in pigs
    Article Snippet: .. Total RNA was prepared from liver samples using TRIzol, prepared for hybridization using the Ambion WT Expression Kit (Life Technologies), following the manufacturer's instructions, except for the input amount of RNA (500 ng input instead of 100 ng) and hybridized in a random order to the Affymetrix Porcine Gene 1.1 ST array (performed by Edinburgh Genomics, University of Edinburgh). ..

    Article Title: Spaceflight Activates Autophagy Programs and the Proteasome in Mouse Liver
    Article Snippet: .. Briefly, RNA was isolated using an RNeasy kit (Qiagen, Germantown, MD, USA) and the Ambion WT expression kit (Thermo Fisher Scientific, Waltham, MA, USA) was employed to prepare mRNA for whole transcriptome microarray analysis using an Affymetrix GeneChip 1.0 ST array (Thermo Fisher Scientific). .. Arrays were scanned using a GeneChip Scanner 3000 7G (Thermo Fisher Scientific) and Command Console Software v. 3.2.3 (Thermo Fisher Scientific) to produce.

    Article Title: Spheres Derived from Lung Adenocarcinoma Pleural Effusions: Molecular Characterization and Tumor Engraftment
    Article Snippet: .. Microarray Analysis Single strand biotinylated cDNA was generated as follows: 100 ng of total RNA were subjected to two cycles of cDNA synthesis with the Ambion WT expression Kit (Applera). ..

    Article Title: Context-dependent anticodon recognition by class I lysyl-tRNA synthetases
    Article Snippet: .. Expand High Fidelity PCR system was from Roche, pBAD-TOPO cloning and expression kit was from Invitrogen, Ni-NTA matrix was from Qiagen (Chatsworth, CA), and L(+) arabinose was from Sigma. .. Mature Escherichia coli tRNALys was purchased from Sigma.

    Article Title: Interhemispheric gene expression differences in the cerebral cortex of humans and macaque monkeys
    Article Snippet: .. Hybridization targets were prepared with an Ambion WT Expression Kit (Applied Biosystems/Ambion, Austin, TX) and Affymetrix GeneChip WT Terminal Labeling Kit from total RNA, hybridized to individual GeneChip® Human Gene 2.0 ST arrays in Affymetrix GeneChip® hybridization oven 645. .. Microarrays were washed in Affymetrix GeneChip® Fluidics Station 450 and scanned with Affymetrix GeneChip® Scanner 7G according to standard Affymetrix GeneChip® hybridization, wash, and stain protocols (Affymetrix, Santa Clara, CA).

    Article Title: Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats
    Article Snippet: .. Affymetrix GeneChip Rat Gene 1.0 ST Array System was used for the microarray analysis following the standard protocol: 100 ng RNA was amplified with Ambion WT Expression Kit (Applied Biosystems), 5.5 µg single-stranded cDNA was labeled and fragmented with GeneChip WT Terminal Labeling and Hybridization (Affymetrix) and hybridized on the chip according to the manufacturer procedure. .. Gene expression determined by real time PCR Total RNA was extracted from liver using Trizol reagent (Invitrogen), and cDNA was prepared and analyzed by real-time PCR testing using QuantiTect SYBR Green reagents (Qiagen, Inc.) on an Opticon continuous fluorescence detector (MJ Research).

    Polymerase Chain Reaction:

    Article Title: Context-dependent anticodon recognition by class I lysyl-tRNA synthetases
    Article Snippet: .. Expand High Fidelity PCR system was from Roche, pBAD-TOPO cloning and expression kit was from Invitrogen, Ni-NTA matrix was from Qiagen (Chatsworth, CA), and L(+) arabinose was from Sigma. .. Mature Escherichia coli tRNALys was purchased from Sigma.

    Chromatin Immunoprecipitation:

    Article Title: Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats
    Article Snippet: .. Affymetrix GeneChip Rat Gene 1.0 ST Array System was used for the microarray analysis following the standard protocol: 100 ng RNA was amplified with Ambion WT Expression Kit (Applied Biosystems), 5.5 µg single-stranded cDNA was labeled and fragmented with GeneChip WT Terminal Labeling and Hybridization (Affymetrix) and hybridized on the chip according to the manufacturer procedure. .. Gene expression determined by real time PCR Total RNA was extracted from liver using Trizol reagent (Invitrogen), and cDNA was prepared and analyzed by real-time PCR testing using QuantiTect SYBR Green reagents (Qiagen, Inc.) on an Opticon continuous fluorescence detector (MJ Research).

    Hybridization:

    Article Title: Macrophage colony-stimulating factor (CSF1) controls monocyte production and maturation and the steady-state size of the liver in pigs
    Article Snippet: .. Total RNA was prepared from liver samples using TRIzol, prepared for hybridization using the Ambion WT Expression Kit (Life Technologies), following the manufacturer's instructions, except for the input amount of RNA (500 ng input instead of 100 ng) and hybridized in a random order to the Affymetrix Porcine Gene 1.1 ST array (performed by Edinburgh Genomics, University of Edinburgh). ..

    Article Title: Interhemispheric gene expression differences in the cerebral cortex of humans and macaque monkeys
    Article Snippet: .. Hybridization targets were prepared with an Ambion WT Expression Kit (Applied Biosystems/Ambion, Austin, TX) and Affymetrix GeneChip WT Terminal Labeling Kit from total RNA, hybridized to individual GeneChip® Human Gene 2.0 ST arrays in Affymetrix GeneChip® hybridization oven 645. .. Microarrays were washed in Affymetrix GeneChip® Fluidics Station 450 and scanned with Affymetrix GeneChip® Scanner 7G according to standard Affymetrix GeneChip® hybridization, wash, and stain protocols (Affymetrix, Santa Clara, CA).

    Article Title: Fumaric Acid Esters Can Block Pro-Inflammatory Actions of Human CRP and Ameliorate Metabolic Disturbances in Transgenic Spontaneously Hypertensive Rats
    Article Snippet: .. Affymetrix GeneChip Rat Gene 1.0 ST Array System was used for the microarray analysis following the standard protocol: 100 ng RNA was amplified with Ambion WT Expression Kit (Applied Biosystems), 5.5 µg single-stranded cDNA was labeled and fragmented with GeneChip WT Terminal Labeling and Hybridization (Affymetrix) and hybridized on the chip according to the manufacturer procedure. .. Gene expression determined by real time PCR Total RNA was extracted from liver using Trizol reagent (Invitrogen), and cDNA was prepared and analyzed by real-time PCR testing using QuantiTect SYBR Green reagents (Qiagen, Inc.) on an Opticon continuous fluorescence detector (MJ Research).

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  • 99
    Thermo Fisher wt expression kit
    Wt Expression Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1019 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/wt expression kit/product/Thermo Fisher
    Average 99 stars, based on 1019 article reviews
    Price from $9.99 to $1999.99
    wt expression kit - by Bioz Stars, 2021-01
    99/100 stars
      Buy from Supplier

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