non targeting dsrna  (Thermo Fisher)


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    Structured Review

    Thermo Fisher non targeting dsrna
    Non Targeting Dsrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/non targeting dsrna/product/Thermo Fisher
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    non targeting dsrna - by Bioz Stars, 2020-07
    88/100 stars

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    Sequencing:

    Article Title: Myosin X Regulates Sealing Zone Patterning in Osteoclasts through Linkage of Podosomes and Microtubules *
    Article Snippet: .. For all experiments, a non-targeting dsRNA from Ambion (Austin, TX) was used as a negative control, and siRNAs homologous to siRNAs 73578 and 73762 but containing three point mutations in the middle of the sequence also were used as negative controls (co73578, co73762). ..

    Negative Control:

    Article Title: Myosin X Regulates Sealing Zone Patterning in Osteoclasts through Linkage of Podosomes and Microtubules *
    Article Snippet: .. For all experiments, a non-targeting dsRNA from Ambion (Austin, TX) was used as a negative control, and siRNAs homologous to siRNAs 73578 and 73762 but containing three point mutations in the middle of the sequence also were used as negative controls (co73578, co73762). ..

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  • 94
    Thermo Fisher dsrna
    Removal of Long <t>dsRNA</t> from IVT mRNA by Cellulose Chromatography (A) Purified, m1Ψ-containing, 1.0-kb-long mRNA was spiked with a 1-kb-long m1Ψ-containing dsRNA at final concentrations of 0.02–20 ng dsRNA/μg mRNA. 100 μg dsRNA-spiked mRNA sample was purified using microcentrifuge spin columns filled with 0.14 g cellulose. The purified mRNA recovered from the flowthrough, unbound fraction and the unpurified input mRNAs (1.0 μg/dot) were analyzed on dot blot. (B) The indicated amounts of 1 kb dsRNA containing m1Ψ were subjected to cellulose purification using microcentrifuge spin columns filled with 0.14 g cellulose. Aliquots of the dsRNA bound to the cellulose or recovered from the flowthrough fractions (unbound) and the input dsRNA were analyzed on dot blots using J2 dsRNA-specific mAb. Aliquots of the corresponding dsRNA samples were also analyzed in non-denaturing 1.4% agarose gel. The <t>GelRed-stained</t> samples were visualized by UV illumination.
    Dsrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 127 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dsrna/product/Thermo Fisher
    Average 94 stars, based on 127 article reviews
    Price from $9.99 to $1999.99
    dsrna - by Bioz Stars, 2020-07
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    90
    Thermo Fisher dsred dsrna
    Expression level of APN1 in the cells transfected with <t>DsRed</t> or HzAPN1 <t>dsRNA.</t> For DsRed (dsRNA control) and HzAPN1 dsRNA, three independent transfections were conducted for each cell line and the protein extracts from each of the three transfections were analyzed by Western blot. The lower panel picture of APN1 and β-actin bands is a representative of the three Western blots. The average expression of HzAPN1 or SfAPN1 (Sf9 cells) relative to that of β-actin calculated by Image J quantification of the three Western blots is displayed in the upper panel. Each error bar represents the standard error of the mean from three transfection replicates. Significant differences in relative expression of HzAPN1 or SfAPN1 (Sf9 cells) between DsRed and HzAPN1 dsRNA transfected cells of each cell line are indicated with an asterisk (*) ( P
    Dsred Dsrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 1 article reviews
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    92
    Thermo Fisher dsrna synthesis
    Real-time quantitative PCR analysis of ACBP (a), FABP10 (b), CPT1 (c), and ACC (d) transcript expressions in the hepatopancreas of M. nipponense injected with either <t>MnSR-BI</t> or GFP <t>dsRNA</t> at 48 h. Bars indicate mean ± SD ( n = 3). ∗ P
    Dsrna Synthesis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dsrna synthesis/product/Thermo Fisher
    Average 92 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    dsrna synthesis - by Bioz Stars, 2020-07
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    89
    Thermo Fisher control dsrna
    Approximately 700 bp DNA sequence upstream of tick antifreeze gene is sufficient to drive iafgp gene expression. ( A ) Schematic representation of the iafgp promoter construct used to analyze promoter activity in tick cells. The schematics is not drawn to scale. ~700 bp DNA sequence (position corresponding to 17757–18463 in GenBank acc. no. DS704943) upstream of iafgp gene containing all tested binding sites (AP-1, HSF-1, TATA-region) was cloned into promoterless <t>pGLuc</t> (pGLuc-P iafgp ) vector. The empty pGLuc vector was used as control. These constructs were transfected into tick cells and luciferase assay was performed. ( B ) Luciferase activity measurements determined from culture supernatants from uninnfected (UI) or A . phagocytophilum -infected (I) ISE6 cells transfected with pGLuc-P iafgp or pGLuc constructs is shown. ( C ) Levels of luciferase transcripts in uninfected (UI) or A . phagocytophilum -infected (I) ISE6 cells transfected with pGLuc-P iafgp or pGLuc constructs is shown. The levels of luciferase transcripts were normalized to the levels of beta-actin transcripts. Open or dark grey circles represents uninfected (UI) or A . phagocytophilum -infected (I) cells transfected with pGluc alone, respectively. Light grey or black circles represents uninfected (UI) or A . phagocytophilum -infected (I) cells transfected with pGLuc-P iafgp , respectively. ( D ) Luciferase activity measurements determined from culture supernatants from A . phagocytophilum -infected ISE6 cells treated with <t>mock-dsRNA</t> or ap-1- dsRNA and transfected with pGLuc-P iafgp constructs is shown. In panels B and D, each circle represents luciferase activity measurement from supernatants collected from one culture well. In panel C, each circle indicates sample generated from cells from one culture well. Statistical analysis was performed using Student’s t test and P value less than 0.05 was considered significant in panels B–D.
    Control Dsrna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 89 stars, based on 5 article reviews
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    Image Search Results


    Removal of Long dsRNA from IVT mRNA by Cellulose Chromatography (A) Purified, m1Ψ-containing, 1.0-kb-long mRNA was spiked with a 1-kb-long m1Ψ-containing dsRNA at final concentrations of 0.02–20 ng dsRNA/μg mRNA. 100 μg dsRNA-spiked mRNA sample was purified using microcentrifuge spin columns filled with 0.14 g cellulose. The purified mRNA recovered from the flowthrough, unbound fraction and the unpurified input mRNAs (1.0 μg/dot) were analyzed on dot blot. (B) The indicated amounts of 1 kb dsRNA containing m1Ψ were subjected to cellulose purification using microcentrifuge spin columns filled with 0.14 g cellulose. Aliquots of the dsRNA bound to the cellulose or recovered from the flowthrough fractions (unbound) and the input dsRNA were analyzed on dot blots using J2 dsRNA-specific mAb. Aliquots of the corresponding dsRNA samples were also analyzed in non-denaturing 1.4% agarose gel. The GelRed-stained samples were visualized by UV illumination.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Facile Method for the Removal of dsRNA Contaminant from In Vitro-Transcribed mRNA

    doi: 10.1016/j.omtn.2019.02.018

    Figure Lengend Snippet: Removal of Long dsRNA from IVT mRNA by Cellulose Chromatography (A) Purified, m1Ψ-containing, 1.0-kb-long mRNA was spiked with a 1-kb-long m1Ψ-containing dsRNA at final concentrations of 0.02–20 ng dsRNA/μg mRNA. 100 μg dsRNA-spiked mRNA sample was purified using microcentrifuge spin columns filled with 0.14 g cellulose. The purified mRNA recovered from the flowthrough, unbound fraction and the unpurified input mRNAs (1.0 μg/dot) were analyzed on dot blot. (B) The indicated amounts of 1 kb dsRNA containing m1Ψ were subjected to cellulose purification using microcentrifuge spin columns filled with 0.14 g cellulose. Aliquots of the dsRNA bound to the cellulose or recovered from the flowthrough fractions (unbound) and the input dsRNA were analyzed on dot blots using J2 dsRNA-specific mAb. Aliquots of the corresponding dsRNA samples were also analyzed in non-denaturing 1.4% agarose gel. The GelRed-stained samples were visualized by UV illumination.

    Article Snippet: To visualize the dsRNA, the agarose gel contained 0.005% (v/v) GelRed, while the polyacrylamide gel was stained post-run with SYBR Gold (Thermo Fisher Scientific) diluted 1:10,000 in Tris/acetic acid/EDTA (TAE) buffer.

    Techniques: Chromatography, Purification, Dot Blot, Agarose Gel Electrophoresis, Staining

    Evaluating the Purity of IVT mRNA following Cellulose Chromatography (A) Uridine (U)- or 1-methylpseudouridine (m1Ψ)-containing, 2.2-kb-long IVT mRNA present in 16% ethanol-containing buffer was purified using cellulose-filled microcentrifuge spin columns. A dot blot loaded with aliquots of mRNA was analyzed for the presence of dsRNA contaminants with J2 mAb and reprobed with complementary oligodeoxynucleotide (ODN). The corresponding mRNA fractions were analyzed by electrophoresis on a 1.4% agarose gel. The GelRed-stained RNAs were visualized by UV illumination. (B) In vitro -transcribed mRNA containing m1Ψ was purified repeatedly in three consecutive cycles using cellulose-filled microcentrifuge spin columns. Aliquots of mRNA were analyzed in dot blot with J2 dsRNA-specific mAb and S9.6 mAb specific for RNA:DNA hybrids. The mRNA fractions separated in 1.4% agarose gel were stained with GelRed and visualized by UV illumination. (C) Linearized plasmid and U-containing dsRNA were mixed in 16% ethanol-containing buffer and then subjected to cellulose chromatography. DNA recovered from the unbound fraction and dsRNA eluted from the cellulose-bound fraction were separated on 1% agarose gel, then stained with GelRed and visualized by UV illumination. M, DNA molecular weight marker.

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: A Facile Method for the Removal of dsRNA Contaminant from In Vitro-Transcribed mRNA

    doi: 10.1016/j.omtn.2019.02.018

    Figure Lengend Snippet: Evaluating the Purity of IVT mRNA following Cellulose Chromatography (A) Uridine (U)- or 1-methylpseudouridine (m1Ψ)-containing, 2.2-kb-long IVT mRNA present in 16% ethanol-containing buffer was purified using cellulose-filled microcentrifuge spin columns. A dot blot loaded with aliquots of mRNA was analyzed for the presence of dsRNA contaminants with J2 mAb and reprobed with complementary oligodeoxynucleotide (ODN). The corresponding mRNA fractions were analyzed by electrophoresis on a 1.4% agarose gel. The GelRed-stained RNAs were visualized by UV illumination. (B) In vitro -transcribed mRNA containing m1Ψ was purified repeatedly in three consecutive cycles using cellulose-filled microcentrifuge spin columns. Aliquots of mRNA were analyzed in dot blot with J2 dsRNA-specific mAb and S9.6 mAb specific for RNA:DNA hybrids. The mRNA fractions separated in 1.4% agarose gel were stained with GelRed and visualized by UV illumination. (C) Linearized plasmid and U-containing dsRNA were mixed in 16% ethanol-containing buffer and then subjected to cellulose chromatography. DNA recovered from the unbound fraction and dsRNA eluted from the cellulose-bound fraction were separated on 1% agarose gel, then stained with GelRed and visualized by UV illumination. M, DNA molecular weight marker.

    Article Snippet: To visualize the dsRNA, the agarose gel contained 0.005% (v/v) GelRed, while the polyacrylamide gel was stained post-run with SYBR Gold (Thermo Fisher Scientific) diluted 1:10,000 in Tris/acetic acid/EDTA (TAE) buffer.

    Techniques: Chromatography, Purification, Dot Blot, Electrophoresis, Agarose Gel Electrophoresis, Staining, In Vitro, Plasmid Preparation, Molecular Weight, Marker

    Expression level of APN1 in the cells transfected with DsRed or HzAPN1 dsRNA. For DsRed (dsRNA control) and HzAPN1 dsRNA, three independent transfections were conducted for each cell line and the protein extracts from each of the three transfections were analyzed by Western blot. The lower panel picture of APN1 and β-actin bands is a representative of the three Western blots. The average expression of HzAPN1 or SfAPN1 (Sf9 cells) relative to that of β-actin calculated by Image J quantification of the three Western blots is displayed in the upper panel. Each error bar represents the standard error of the mean from three transfection replicates. Significant differences in relative expression of HzAPN1 or SfAPN1 (Sf9 cells) between DsRed and HzAPN1 dsRNA transfected cells of each cell line are indicated with an asterisk (*) ( P

    Journal: Scientific Reports

    Article Title: APN1 is a functional receptor of Cry1Ac but not Cry2Ab in Helicoverpa zea

    doi: 10.1038/srep19179

    Figure Lengend Snippet: Expression level of APN1 in the cells transfected with DsRed or HzAPN1 dsRNA. For DsRed (dsRNA control) and HzAPN1 dsRNA, three independent transfections were conducted for each cell line and the protein extracts from each of the three transfections were analyzed by Western blot. The lower panel picture of APN1 and β-actin bands is a representative of the three Western blots. The average expression of HzAPN1 or SfAPN1 (Sf9 cells) relative to that of β-actin calculated by Image J quantification of the three Western blots is displayed in the upper panel. Each error bar represents the standard error of the mean from three transfection replicates. Significant differences in relative expression of HzAPN1 or SfAPN1 (Sf9 cells) between DsRed and HzAPN1 dsRNA transfected cells of each cell line are indicated with an asterisk (*) ( P

    Article Snippet: Six μg protein of the larval midgut BBMV and/or the protein extract of each cell line transfected with pAc, pAc-HzAPN1, DsRed dsRNA, or HzAPN1 dsRNA were separated on 10% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) gel and electroblotted to a polyvinylidene difluoride (PVDF) membrane (Thermo scientific) in the transfer buffer (25 mM Tris, 192 mM glycine, 10% methanol, pH 8.3).

    Techniques: Expressing, Transfection, Western Blot

    Real-time quantitative PCR analysis of ACBP (a), FABP10 (b), CPT1 (c), and ACC (d) transcript expressions in the hepatopancreas of M. nipponense injected with either MnSR-BI or GFP dsRNA at 48 h. Bars indicate mean ± SD ( n = 3). ∗ P

    Journal: International Journal of Genomics

    Article Title: Scavenger Receptor Class B, Type I, a CD36 Related Protein in Macrobrachium nipponense: Characterization, RNA Interference, and Expression Analysis with Different Dietary Lipid Sources

    doi: 10.1155/2016/6325927

    Figure Lengend Snippet: Real-time quantitative PCR analysis of ACBP (a), FABP10 (b), CPT1 (c), and ACC (d) transcript expressions in the hepatopancreas of M. nipponense injected with either MnSR-BI or GFP dsRNA at 48 h. Bars indicate mean ± SD ( n = 3). ∗ P

    Article Snippet: 2.5. dsRNA Synthesis and Injection Either MnSR-BI dsRNA or green fluorescent protein (GFP) was synthesized in vitro with a Transcript Aid™ T7 High Yield Transcription Kit (Thermo Scientific, Inc., Massachusetts, USA) according to the manufacturer's instructions.

    Techniques: Real-time Polymerase Chain Reaction, Injection

    Real-time quantitative PCR analysis of the MnSR-BI transcript expression in the hepatopancreas of M. nipponense injected with either MnSR-BI or GFP dsRNA. Samples were obtained 48 h (a) and 96 h (b) after dsRNA injection and analyzed by real-time qRT-PCR. Bars indicate mean ± SD ( n = 3). ∗ P

    Journal: International Journal of Genomics

    Article Title: Scavenger Receptor Class B, Type I, a CD36 Related Protein in Macrobrachium nipponense: Characterization, RNA Interference, and Expression Analysis with Different Dietary Lipid Sources

    doi: 10.1155/2016/6325927

    Figure Lengend Snippet: Real-time quantitative PCR analysis of the MnSR-BI transcript expression in the hepatopancreas of M. nipponense injected with either MnSR-BI or GFP dsRNA. Samples were obtained 48 h (a) and 96 h (b) after dsRNA injection and analyzed by real-time qRT-PCR. Bars indicate mean ± SD ( n = 3). ∗ P

    Article Snippet: 2.5. dsRNA Synthesis and Injection Either MnSR-BI dsRNA or green fluorescent protein (GFP) was synthesized in vitro with a Transcript Aid™ T7 High Yield Transcription Kit (Thermo Scientific, Inc., Massachusetts, USA) according to the manufacturer's instructions.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Injection, Quantitative RT-PCR

    Approximately 700 bp DNA sequence upstream of tick antifreeze gene is sufficient to drive iafgp gene expression. ( A ) Schematic representation of the iafgp promoter construct used to analyze promoter activity in tick cells. The schematics is not drawn to scale. ~700 bp DNA sequence (position corresponding to 17757–18463 in GenBank acc. no. DS704943) upstream of iafgp gene containing all tested binding sites (AP-1, HSF-1, TATA-region) was cloned into promoterless pGLuc (pGLuc-P iafgp ) vector. The empty pGLuc vector was used as control. These constructs were transfected into tick cells and luciferase assay was performed. ( B ) Luciferase activity measurements determined from culture supernatants from uninnfected (UI) or A . phagocytophilum -infected (I) ISE6 cells transfected with pGLuc-P iafgp or pGLuc constructs is shown. ( C ) Levels of luciferase transcripts in uninfected (UI) or A . phagocytophilum -infected (I) ISE6 cells transfected with pGLuc-P iafgp or pGLuc constructs is shown. The levels of luciferase transcripts were normalized to the levels of beta-actin transcripts. Open or dark grey circles represents uninfected (UI) or A . phagocytophilum -infected (I) cells transfected with pGluc alone, respectively. Light grey or black circles represents uninfected (UI) or A . phagocytophilum -infected (I) cells transfected with pGLuc-P iafgp , respectively. ( D ) Luciferase activity measurements determined from culture supernatants from A . phagocytophilum -infected ISE6 cells treated with mock-dsRNA or ap-1- dsRNA and transfected with pGLuc-P iafgp constructs is shown. In panels B and D, each circle represents luciferase activity measurement from supernatants collected from one culture well. In panel C, each circle indicates sample generated from cells from one culture well. Statistical analysis was performed using Student’s t test and P value less than 0.05 was considered significant in panels B–D.

    Journal: Scientific Reports

    Article Title: Arthropod transcriptional activator protein-1 (AP-1) aids tick-rickettsial pathogen survival in the cold

    doi: 10.1038/s41598-018-29654-6

    Figure Lengend Snippet: Approximately 700 bp DNA sequence upstream of tick antifreeze gene is sufficient to drive iafgp gene expression. ( A ) Schematic representation of the iafgp promoter construct used to analyze promoter activity in tick cells. The schematics is not drawn to scale. ~700 bp DNA sequence (position corresponding to 17757–18463 in GenBank acc. no. DS704943) upstream of iafgp gene containing all tested binding sites (AP-1, HSF-1, TATA-region) was cloned into promoterless pGLuc (pGLuc-P iafgp ) vector. The empty pGLuc vector was used as control. These constructs were transfected into tick cells and luciferase assay was performed. ( B ) Luciferase activity measurements determined from culture supernatants from uninnfected (UI) or A . phagocytophilum -infected (I) ISE6 cells transfected with pGLuc-P iafgp or pGLuc constructs is shown. ( C ) Levels of luciferase transcripts in uninfected (UI) or A . phagocytophilum -infected (I) ISE6 cells transfected with pGLuc-P iafgp or pGLuc constructs is shown. The levels of luciferase transcripts were normalized to the levels of beta-actin transcripts. Open or dark grey circles represents uninfected (UI) or A . phagocytophilum -infected (I) cells transfected with pGluc alone, respectively. Light grey or black circles represents uninfected (UI) or A . phagocytophilum -infected (I) cells transfected with pGLuc-P iafgp , respectively. ( D ) Luciferase activity measurements determined from culture supernatants from A . phagocytophilum -infected ISE6 cells treated with mock-dsRNA or ap-1- dsRNA and transfected with pGLuc-P iafgp constructs is shown. In panels B and D, each circle represents luciferase activity measurement from supernatants collected from one culture well. In panel C, each circle indicates sample generated from cells from one culture well. Statistical analysis was performed using Student’s t test and P value less than 0.05 was considered significant in panels B–D.

    Article Snippet: After incubation, 750 ng of ap-1 dsRNA or equal volume of mock solution or control dsRNA or pGLuc/pGLuc-Piafgp constructs were mixed with Lipofectamine 2000 (Thermo Fisher Scientific, USA) reagent and added to the wells.

    Techniques: Sequencing, Expressing, Construct, Activity Assay, Binding Assay, Clone Assay, Plasmid Preparation, Transfection, Luciferase, Infection, Generated