single molecule mrna fish probe  (Thermo Fisher)


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    Structured Review

    Thermo Fisher single molecule mrna fish probe
    Technical controls and the definition of a yellow spot. ( A ) Red and green fluorescence in situ hybridization <t>(FISH)</t> probes were designed to anneal to the same <t>mRNA</t> sequence of an average-sized mRNA (Tb427.10.14550). The signals from the two probes showed a strong overlap. One representative cell is shown; more cells are shown in Supplementary Figure S2 . ( B ) Red and green FISH probes were designed to anneal to adjacent mRNA sequences of the same, average-sized mRNA. The signals from the two probes again showed a strong overlap. One representative cell is shown; more cells are shown in Supplementary Figure S3 . ( C–E ) Trypanosome cells were simultaneously probed with the 5΄ probe (red) and the 3΄ probe (green) specific for the GB4 mRNA (Tb427tmp.160.1200) (details on top left of C). Together, these probes cover the entire 24 645 nucleotides of the GB4 open reading frame: the 5΄ probe binds to the 2256 nucleotides at the 5΄ end and the 3΄ probe to the remaining 22 389 nucleotides. ( C ) One representative Z-stack projection image with merged fluorescence channels is shown. ( D ) The length of 100 yellow GB4 mRNA molecules was measured from Z-stack projection images. In total, 97% are less than 0.5 μm in length (D). ( E ) Example images of very extended GB4 mRNA molecules; the length is indicated on top and the blue line shows how the length measurement was performed (from the middle of the ‘red circle’ to the middle of the most distant ‘green circle’ at the end of the green string). These very long mRNA molecules are rare and never longer than 1.0 μm. ( F ) Trans-probing: dual colour mRNA FISH using probes antisense to two different mRNAs (details on top left). The percentage of red, green and yellow spots was quantified from 152 cells (top right). The average cell had 7.3 ± 2.8 mRNA molecules. A representative image is shown (bottom).
    Single Molecule Mrna Fish Probe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/single molecule mrna fish probe/product/Thermo Fisher
    Average 91 stars, based on 1 article reviews
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    single molecule mrna fish probe - by Bioz Stars, 2020-09
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    1) Product Images from "Simultaneous detection of mRNA transcription and decay intermediates by dual colour single mRNA FISH on subcellular resolution"

    Article Title: Simultaneous detection of mRNA transcription and decay intermediates by dual colour single mRNA FISH on subcellular resolution

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw1245

    Technical controls and the definition of a yellow spot. ( A ) Red and green fluorescence in situ hybridization (FISH) probes were designed to anneal to the same mRNA sequence of an average-sized mRNA (Tb427.10.14550). The signals from the two probes showed a strong overlap. One representative cell is shown; more cells are shown in Supplementary Figure S2 . ( B ) Red and green FISH probes were designed to anneal to adjacent mRNA sequences of the same, average-sized mRNA. The signals from the two probes again showed a strong overlap. One representative cell is shown; more cells are shown in Supplementary Figure S3 . ( C–E ) Trypanosome cells were simultaneously probed with the 5΄ probe (red) and the 3΄ probe (green) specific for the GB4 mRNA (Tb427tmp.160.1200) (details on top left of C). Together, these probes cover the entire 24 645 nucleotides of the GB4 open reading frame: the 5΄ probe binds to the 2256 nucleotides at the 5΄ end and the 3΄ probe to the remaining 22 389 nucleotides. ( C ) One representative Z-stack projection image with merged fluorescence channels is shown. ( D ) The length of 100 yellow GB4 mRNA molecules was measured from Z-stack projection images. In total, 97% are less than 0.5 μm in length (D). ( E ) Example images of very extended GB4 mRNA molecules; the length is indicated on top and the blue line shows how the length measurement was performed (from the middle of the ‘red circle’ to the middle of the most distant ‘green circle’ at the end of the green string). These very long mRNA molecules are rare and never longer than 1.0 μm. ( F ) Trans-probing: dual colour mRNA FISH using probes antisense to two different mRNAs (details on top left). The percentage of red, green and yellow spots was quantified from 152 cells (top right). The average cell had 7.3 ± 2.8 mRNA molecules. A representative image is shown (bottom).
    Figure Legend Snippet: Technical controls and the definition of a yellow spot. ( A ) Red and green fluorescence in situ hybridization (FISH) probes were designed to anneal to the same mRNA sequence of an average-sized mRNA (Tb427.10.14550). The signals from the two probes showed a strong overlap. One representative cell is shown; more cells are shown in Supplementary Figure S2 . ( B ) Red and green FISH probes were designed to anneal to adjacent mRNA sequences of the same, average-sized mRNA. The signals from the two probes again showed a strong overlap. One representative cell is shown; more cells are shown in Supplementary Figure S3 . ( C–E ) Trypanosome cells were simultaneously probed with the 5΄ probe (red) and the 3΄ probe (green) specific for the GB4 mRNA (Tb427tmp.160.1200) (details on top left of C). Together, these probes cover the entire 24 645 nucleotides of the GB4 open reading frame: the 5΄ probe binds to the 2256 nucleotides at the 5΄ end and the 3΄ probe to the remaining 22 389 nucleotides. ( C ) One representative Z-stack projection image with merged fluorescence channels is shown. ( D ) The length of 100 yellow GB4 mRNA molecules was measured from Z-stack projection images. In total, 97% are less than 0.5 μm in length (D). ( E ) Example images of very extended GB4 mRNA molecules; the length is indicated on top and the blue line shows how the length measurement was performed (from the middle of the ‘red circle’ to the middle of the most distant ‘green circle’ at the end of the green string). These very long mRNA molecules are rare and never longer than 1.0 μm. ( F ) Trans-probing: dual colour mRNA FISH using probes antisense to two different mRNAs (details on top left). The percentage of red, green and yellow spots was quantified from 152 cells (top right). The average cell had 7.3 ± 2.8 mRNA molecules. A representative image is shown (bottom).

    Techniques Used: Fluorescence, In Situ Hybridization, Fluorescence In Situ Hybridization, Sequencing

    Related Articles

    Fluorescence In Situ Hybridization:

    Article Title: Simultaneous detection of mRNA transcription and decay intermediates by dual colour single mRNA FISH on subcellular resolution
    Article Snippet: .. Affymetrix® single molecule mRNA FISH probe sets were designed antisense to the 1100 most 5΄ nucleotides (red fluorescence) and the 1100 most 3΄ nucleotides (green fluorescence) of Tb427.01.1740 and Tb427.04.310. ..

    Fluorescence:

    Article Title: Simultaneous detection of mRNA transcription and decay intermediates by dual colour single mRNA FISH on subcellular resolution
    Article Snippet: .. Affymetrix® single molecule mRNA FISH probe sets were designed antisense to the 1100 most 5΄ nucleotides (red fluorescence) and the 1100 most 3΄ nucleotides (green fluorescence) of Tb427.01.1740 and Tb427.04.310. ..

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    Thermo Fisher random cdna synthesis htt mrna lowering
    <t>HTT</t> <t>mRNA</t> and Mutant Htt Protein Lowering in HD71 Patient iPSC-Derived Neuronal Culture (A) HTT mRNA levels relative to the mean expression in control-treated cells (AAV5-GFP at a MOI of 10 7 ) were determined by gene-specific TaqMan qPCR. (B) Ultra-sensitive single molecule counting assay with 2B7 and MAB2166 antibodies to quantify human Htt protein (both wild-type and mutant), relative to AAV5-GFP at a MOI of 10 7 . Data were evaluated using a one-way ANOVA and corrected using a Bonferonni test. **p
    Random Cdna Synthesis Htt Mrna Lowering, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 93 stars, based on 1 article reviews
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    random cdna synthesis htt mrna lowering - by Bioz Stars, 2020-09
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    HTT mRNA and Mutant Htt Protein Lowering in HD71 Patient iPSC-Derived Neuronal Culture (A) HTT mRNA levels relative to the mean expression in control-treated cells (AAV5-GFP at a MOI of 10 7 ) were determined by gene-specific TaqMan qPCR. (B) Ultra-sensitive single molecule counting assay with 2B7 and MAB2166 antibodies to quantify human Htt protein (both wild-type and mutant), relative to AAV5-GFP at a MOI of 10 7 . Data were evaluated using a one-way ANOVA and corrected using a Bonferonni test. **p

    Journal: Molecular Therapy. Methods & Clinical Development

    Article Title: AAV5-miHTT Lowers Huntingtin mRNA and Protein without Off-Target Effects in Patient-Derived Neuronal Cultures and Astrocytes

    doi: 10.1016/j.omtm.2019.09.010

    Figure Lengend Snippet: HTT mRNA and Mutant Htt Protein Lowering in HD71 Patient iPSC-Derived Neuronal Culture (A) HTT mRNA levels relative to the mean expression in control-treated cells (AAV5-GFP at a MOI of 10 7 ) were determined by gene-specific TaqMan qPCR. (B) Ultra-sensitive single molecule counting assay with 2B7 and MAB2166 antibodies to quantify human Htt protein (both wild-type and mutant), relative to AAV5-GFP at a MOI of 10 7 . Data were evaluated using a one-way ANOVA and corrected using a Bonferonni test. **p

    Article Snippet: Random cDNA Synthesis HTT mRNA lowering was assessed using the DyNAmo cDNA synthesis kit with random hexamer primers (Thermo Fisher Scientific, Loughborough, UK).

    Techniques: Mutagenesis, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Single Molecule Counting