taqman preamp master mix kit  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 97
    Name:
    TaqMan PreAmp Master Mix Kit
    Description:
    Stretch your precious samples into more real time PCR experiments with TaqMan PreAmp Master Mix Kit Applied Biosystems TaqMan PreAmp Master Mix Kit contains TaqMan PreAmp Master Mix and TaqMan Gene Expression Master Mix The two work hand in hand to provide a seamless workflow for preamplification and quantitation of mRNA The TaqMan PreAmp Master Mix Kit lets you • Amplify cDNA targets equally without introducing bias• Analyze mRNA from any precious sample such as laser capture microdissections needle biopsies and formalin fixed paraffin embedded tissues FFPE • Stretch as little as 1 ng of cDNA into 200 real time PCR reactions for gene expression analysis using TaqMan Gene Expression Assays• Analyze up to 100 gene expression targets with minimal hands on timePreserves Equilibrium of Targets Without Pre amplification BiasIn the past preamplification kits and techniques often resulted in uneven amplification of targets and inaccurate or biased data as shown by low correlation coefficients between amplified and unamplified samples TaqMan PreAmp Master Mix amplifies your small starting samples with no bias and provides extremely high correlation between amplified and unamplified cDNA Optimized Workflow and Results TaqMan PreAmp Master Mix and TaqMan Gene Expression Master Mix work in conjunction to provide an optimal workflow for analysis of small starting samples TaqMan Gene Expression Master Mix provides reliable quantification over 9 logs of linear dynamic range and has been validated with TaqMan Assays
    Catalog Number:
    4384267
    Price:
    None
    Applications:
    Enzymes & Master Mixes for Real-Time PCR|Industrial & Applied Science|PCR & Real-Time PCR|PCR-Based Drug Discovery Assays|Pharma & Biopharma|RNAi, Epigenetics & Non-Coding RNA Research|Real Time PCR (qPCR)|Real Time PCR-Based Gene Expression Profiling|Real Time PCR-Based miRNA Analysis|miRNA & Non-Coding RNA Analysis|Target & Lead Identification & Validation|Gene Expression Analysis & Genotyping
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher taqman preamp master mix kit
    Relative levels of mmu-miR-466h and its target genes in fresh and nutrient-depleted media with and without mmu-miR-466h inhibition. (A) Mmu-miR-466h levels. <t>TaqMan</t> microRNA <t>qRT-PCR</t> analysis was used to assess mmu-miR-466h levels at 23.5h in different
    Stretch your precious samples into more real time PCR experiments with TaqMan PreAmp Master Mix Kit Applied Biosystems TaqMan PreAmp Master Mix Kit contains TaqMan PreAmp Master Mix and TaqMan Gene Expression Master Mix The two work hand in hand to provide a seamless workflow for preamplification and quantitation of mRNA The TaqMan PreAmp Master Mix Kit lets you • Amplify cDNA targets equally without introducing bias• Analyze mRNA from any precious sample such as laser capture microdissections needle biopsies and formalin fixed paraffin embedded tissues FFPE • Stretch as little as 1 ng of cDNA into 200 real time PCR reactions for gene expression analysis using TaqMan Gene Expression Assays• Analyze up to 100 gene expression targets with minimal hands on timePreserves Equilibrium of Targets Without Pre amplification BiasIn the past preamplification kits and techniques often resulted in uneven amplification of targets and inaccurate or biased data as shown by low correlation coefficients between amplified and unamplified samples TaqMan PreAmp Master Mix amplifies your small starting samples with no bias and provides extremely high correlation between amplified and unamplified cDNA Optimized Workflow and Results TaqMan PreAmp Master Mix and TaqMan Gene Expression Master Mix work in conjunction to provide an optimal workflow for analysis of small starting samples TaqMan Gene Expression Master Mix provides reliable quantification over 9 logs of linear dynamic range and has been validated with TaqMan Assays
    https://www.bioz.com/result/taqman preamp master mix kit/product/Thermo Fisher
    Average 97 stars, based on 111 article reviews
    Price from $9.99 to $1999.99
    taqman preamp master mix kit - by Bioz Stars, 2020-07
    97/100 stars

    Images

    1) Product Images from "A novel microRNA mmu-mir-466h affects apoptosis regulation in mammalian cells"

    Article Title: A novel microRNA mmu-mir-466h affects apoptosis regulation in mammalian cells

    Journal: Biotechnology and bioengineering

    doi: 10.1002/bit.23092

    Relative levels of mmu-miR-466h and its target genes in fresh and nutrient-depleted media with and without mmu-miR-466h inhibition. (A) Mmu-miR-466h levels. TaqMan microRNA qRT-PCR analysis was used to assess mmu-miR-466h levels at 23.5h in different
    Figure Legend Snippet: Relative levels of mmu-miR-466h and its target genes in fresh and nutrient-depleted media with and without mmu-miR-466h inhibition. (A) Mmu-miR-466h levels. TaqMan microRNA qRT-PCR analysis was used to assess mmu-miR-466h levels at 23.5h in different

    Techniques Used: Inhibition, Quantitative RT-PCR

    qRT-PCR comparison of mmu-miR-466h, mmu-miR-669c, and mRNA levels for mmu-miR-466h predicted targets in fresh and depleted media after 24h. (A) Mmu-miR-466h and mmu-miR-669c levels. The TaqMan microRNA assays were used for both miRs with sno202 and let-7c
    Figure Legend Snippet: qRT-PCR comparison of mmu-miR-466h, mmu-miR-669c, and mRNA levels for mmu-miR-466h predicted targets in fresh and depleted media after 24h. (A) Mmu-miR-466h and mmu-miR-669c levels. The TaqMan microRNA assays were used for both miRs with sno202 and let-7c

    Techniques Used: Quantitative RT-PCR

    2) Product Images from "miR-524-5p suppresses the growth of oncogenic BRAF melanoma by targeting BRAF and ERK2"

    Article Title: miR-524-5p suppresses the growth of oncogenic BRAF melanoma by targeting BRAF and ERK2

    Journal: Oncotarget

    doi:

    The expression of miR-214, miR-433, and miR-524-5p is suppressed in melanoma (A) Left panel, MAPK/ERK signaling was highly activated in the Malme-3M cell line according to Western analysis to detect the protein levels of BRAF and phospho-MEK. Right panel, the relative expression levels of miR-214, miR-433, and miR-524-5p were detected by the TaqMan miRNA expression array (normalized to RNU44 and RNU48) with the ratio of Malme-3 to Malme-3M. (B-D) Box-whisker plots of miR-214, miR-433, and miR-524-5p in melanoma samples. The miRNA expression profiles were obtained from the Gene Expression Omnibus (GEO) accession numbers GSE20994 and GSE31568. We downloaded raw data and used statistical analysis to obtain the mean and FDR values. The fold change was evaluated from the mean of melanoma samples versus normal.
    Figure Legend Snippet: The expression of miR-214, miR-433, and miR-524-5p is suppressed in melanoma (A) Left panel, MAPK/ERK signaling was highly activated in the Malme-3M cell line according to Western analysis to detect the protein levels of BRAF and phospho-MEK. Right panel, the relative expression levels of miR-214, miR-433, and miR-524-5p were detected by the TaqMan miRNA expression array (normalized to RNU44 and RNU48) with the ratio of Malme-3 to Malme-3M. (B-D) Box-whisker plots of miR-214, miR-433, and miR-524-5p in melanoma samples. The miRNA expression profiles were obtained from the Gene Expression Omnibus (GEO) accession numbers GSE20994 and GSE31568. We downloaded raw data and used statistical analysis to obtain the mean and FDR values. The fold change was evaluated from the mean of melanoma samples versus normal.

    Techniques Used: Expressing, Western Blot, Whisker Assay

    3) Product Images from "Functional mechanisms underlying pleiotropic risk alleles at the 19p13.1 breast–ovarian cancer susceptibility locus"

    Article Title: Functional mechanisms underlying pleiotropic risk alleles at the 19p13.1 breast–ovarian cancer susceptibility locus

    Journal: Nature Communications

    doi: 10.1038/ncomms12675

    Effects of deletion of the putative enhancer containing the rs56069439 risk SNP in breast and ovarian epithelial cells. ( a ) Illustration of the 57 bp region in an intron of ANKLE1 containing rs56069439; H3K4me1 marks overlapped rs56069439 in ovarian, fallopian and breast cells. Location of the two guide RNAs (gRNAs) used to create the stable Δrs56069439 deletion by CRISPR/Cas9 genome editing, cutting sites are indicated with the green arrow. PAM, protospacer adjacent motif. ( b ) PCR analysis of targeted region in representative MCF10A (breast) epithelial cell clones. Control clones were transfected with the vector backbone only. ( c ) Verification of deletions by Sanger sequencing, and alignment to the genome using BLAT. ( d ) Gene expression analysis using TaqMan probes showing downregulation of ANKLE1 was associated with deletion of a region containing rs56069439.
    Figure Legend Snippet: Effects of deletion of the putative enhancer containing the rs56069439 risk SNP in breast and ovarian epithelial cells. ( a ) Illustration of the 57 bp region in an intron of ANKLE1 containing rs56069439; H3K4me1 marks overlapped rs56069439 in ovarian, fallopian and breast cells. Location of the two guide RNAs (gRNAs) used to create the stable Δrs56069439 deletion by CRISPR/Cas9 genome editing, cutting sites are indicated with the green arrow. PAM, protospacer adjacent motif. ( b ) PCR analysis of targeted region in representative MCF10A (breast) epithelial cell clones. Control clones were transfected with the vector backbone only. ( c ) Verification of deletions by Sanger sequencing, and alignment to the genome using BLAT. ( d ) Gene expression analysis using TaqMan probes showing downregulation of ANKLE1 was associated with deletion of a region containing rs56069439.

    Techniques Used: CRISPR, Polymerase Chain Reaction, Clone Assay, Transfection, Plasmid Preparation, Sequencing, Expressing

    Related Articles

    Amplification:

    Article Title: Functional mechanisms underlying pleiotropic risk alleles at the 19p13.1 breast–ovarian cancer susceptibility locus
    Article Snippet: .. The cDNA was diluted to 10 ng μl−1 and 12.5 ng was used in target specific amplification before real-time PCR using TaqMan PreAmp Master Mix Kit (Applied Biosystems) following Fluidigm's Specific Target Amplification Protocol. .. 1.25 μl of the 25 μl pre-amplified cDNA was added to each chip.

    Synthesized:

    Article Title: miR-524-5p suppresses the growth of oncogenic BRAF melanoma by targeting BRAF and ERK2
    Article Snippet: .. qRT–PCR Total RNA was extracted from the cells (or tumor tissues) using mirVana™ miRNA Isolation Kit (Ambion) according to the manufacturer's protocol. miR-524-5p and RNU48 cDNA were synthesized by TaqMan PreAmp Master Mix Kit and TaqMan MicroRNA Assays (Applied Biosystems) according to the manufacturer's instructions to quantify microRNA expression. .. Relative expression of target microRNAs was calculated using the ΔΔ Ct method and normalized to RNU48.

    Article Title: Induction of Immune Tolerance to a Therapeutic Protein by Intrathymic Gene Delivery
    Article Snippet: .. Total RNA concentrations were quantified by A260 (NanoDrop ND-1000 Spectrophotometer; NanoDrop Technologies, Wilmington, DE). cDNA was synthesized and preamplificated (except for hGAA in liver) using the TaqMan PreAmplification master mix kit (Applied Biosystems), amplifying 15 cycles for hGAA in thymus (without preamplification thymic hGAA mRNA copies were below the detection limit of 100 copies/125 ng total RNA; for increased accuracy, the resulting copy numbers are expressed as a percentage of copies found after intrathymic delivery), and 14 cycles for other genes. .. A pooled TaqMan assay for the preamplification step contained TaqMan Gene Expression Assay (Applied Biosystems) for various genes at 0.2× dilution as recommended by the kit, with the exception of a 0.002× dilution for mouse actin.

    Isolation:

    Article Title: miR-524-5p suppresses the growth of oncogenic BRAF melanoma by targeting BRAF and ERK2
    Article Snippet: .. qRT–PCR Total RNA was extracted from the cells (or tumor tissues) using mirVana™ miRNA Isolation Kit (Ambion) according to the manufacturer's protocol. miR-524-5p and RNU48 cDNA were synthesized by TaqMan PreAmp Master Mix Kit and TaqMan MicroRNA Assays (Applied Biosystems) according to the manufacturer's instructions to quantify microRNA expression. .. Relative expression of target microRNAs was calculated using the ΔΔ Ct method and normalized to RNU48.

    Spectrophotometry:

    Article Title: Induction of Immune Tolerance to a Therapeutic Protein by Intrathymic Gene Delivery
    Article Snippet: .. Total RNA concentrations were quantified by A260 (NanoDrop ND-1000 Spectrophotometer; NanoDrop Technologies, Wilmington, DE). cDNA was synthesized and preamplificated (except for hGAA in liver) using the TaqMan PreAmplification master mix kit (Applied Biosystems), amplifying 15 cycles for hGAA in thymus (without preamplification thymic hGAA mRNA copies were below the detection limit of 100 copies/125 ng total RNA; for increased accuracy, the resulting copy numbers are expressed as a percentage of copies found after intrathymic delivery), and 14 cycles for other genes. .. A pooled TaqMan assay for the preamplification step contained TaqMan Gene Expression Assay (Applied Biosystems) for various genes at 0.2× dilution as recommended by the kit, with the exception of a 0.002× dilution for mouse actin.

    Real-time Polymerase Chain Reaction:

    Article Title: Prognostic Impact of Modulators of G proteins in Circulating Tumor Cells from Patients with Metastatic Colorectal Cancer
    Article Snippet: .. To optimize target detection, samples were first preamplified (PreAmp Master Mix kit, Life Technologies). mRNA levels of CD45, CCDC88A, CCDC88C, NUCB1, NUCB1, S100A4 and MACC1 genes were quantified by quantitative Real-Time PCR using hydrolysis probes chemistry (Life Technologies) in a StepOne plus thermocycler (Life Technologies). ..

    Article Title: Functional mechanisms underlying pleiotropic risk alleles at the 19p13.1 breast–ovarian cancer susceptibility locus
    Article Snippet: .. The cDNA was diluted to 10 ng μl−1 and 12.5 ng was used in target specific amplification before real-time PCR using TaqMan PreAmp Master Mix Kit (Applied Biosystems) following Fluidigm's Specific Target Amplification Protocol. .. 1.25 μl of the 25 μl pre-amplified cDNA was added to each chip.

    Quantitative RT-PCR:

    Article Title: microRNA Expression in Sentinel Nodes from Progressing Melanoma Patients Identifies Networks Associated with Dysfunctional Immune Response
    Article Snippet: .. Gene and miR expression levels were evaluated by qRT-PCR using Thermo Fisher Scientific reagents (Waltham, MA, USA) with the exception of miR-574-5p. microRNA Reverse Transcription Kit (Thermo Fisher), with the associated RT primer pool, or High-Capacity cDNA Archive Kit (Thermo Fisher) were used to retrotranscribe miRs and mRNAs, respectively; TaqMan PreAmp Master Mix (Thermo Fisher) with the associated pre-amplification primer pool was used for pre-amplifications. miR-574-5p expression was assessed by Qiagen assays (Qiagen, Hilden, Germany), after retrotranscription by miScript II RT Kit (Thermo Fisher) and pre-amplification by miScript PreAMP PCR Kit (Thermo Fisher) using as internal reference U6 snRNA (Qiagen, Hilden, Germany). .. TNFRSF8 , TNF Receptor Superfamily Member 8 (Hs00174277_m1) and Beta Actin (4326315E) TaqMan Gene Expression assays were used.

    Article Title: miR-524-5p suppresses the growth of oncogenic BRAF melanoma by targeting BRAF and ERK2
    Article Snippet: .. qRT–PCR Total RNA was extracted from the cells (or tumor tissues) using mirVana™ miRNA Isolation Kit (Ambion) according to the manufacturer's protocol. miR-524-5p and RNU48 cDNA were synthesized by TaqMan PreAmp Master Mix Kit and TaqMan MicroRNA Assays (Applied Biosystems) according to the manufacturer's instructions to quantify microRNA expression. .. Relative expression of target microRNAs was calculated using the ΔΔ Ct method and normalized to RNU48.

    Article Title: A novel microRNA mmu-mir-466h affects apoptosis regulation in mammalian cells
    Article Snippet: .. The preamplification (10 cycles) was done using Applied Systems TaqMan® PreAmp Kit (Part No. 4384267) in PCR Thermal cycler (Applied Biosystems) after miRNA reverse transcription and before qRT-PCR reads. .. TaqMan® PreAmp Master Mix preamplifies small amounts of cDNA without introducing amplification bias to the sample and provides a very high correlation coefficients between amplified and unamplified cDNA ( ). qRT-PCR measurements were performed in quadruplicates.

    Expressing:

    Article Title: microRNA Expression in Sentinel Nodes from Progressing Melanoma Patients Identifies Networks Associated with Dysfunctional Immune Response
    Article Snippet: .. Gene and miR expression levels were evaluated by qRT-PCR using Thermo Fisher Scientific reagents (Waltham, MA, USA) with the exception of miR-574-5p. microRNA Reverse Transcription Kit (Thermo Fisher), with the associated RT primer pool, or High-Capacity cDNA Archive Kit (Thermo Fisher) were used to retrotranscribe miRs and mRNAs, respectively; TaqMan PreAmp Master Mix (Thermo Fisher) with the associated pre-amplification primer pool was used for pre-amplifications. miR-574-5p expression was assessed by Qiagen assays (Qiagen, Hilden, Germany), after retrotranscription by miScript II RT Kit (Thermo Fisher) and pre-amplification by miScript PreAMP PCR Kit (Thermo Fisher) using as internal reference U6 snRNA (Qiagen, Hilden, Germany). .. TNFRSF8 , TNF Receptor Superfamily Member 8 (Hs00174277_m1) and Beta Actin (4326315E) TaqMan Gene Expression assays were used.

    Article Title: miR-524-5p suppresses the growth of oncogenic BRAF melanoma by targeting BRAF and ERK2
    Article Snippet: .. qRT–PCR Total RNA was extracted from the cells (or tumor tissues) using mirVana™ miRNA Isolation Kit (Ambion) according to the manufacturer's protocol. miR-524-5p and RNU48 cDNA were synthesized by TaqMan PreAmp Master Mix Kit and TaqMan MicroRNA Assays (Applied Biosystems) according to the manufacturer's instructions to quantify microRNA expression. .. Relative expression of target microRNAs was calculated using the ΔΔ Ct method and normalized to RNU48.

    Polymerase Chain Reaction:

    Article Title: microRNA Expression in Sentinel Nodes from Progressing Melanoma Patients Identifies Networks Associated with Dysfunctional Immune Response
    Article Snippet: .. Gene and miR expression levels were evaluated by qRT-PCR using Thermo Fisher Scientific reagents (Waltham, MA, USA) with the exception of miR-574-5p. microRNA Reverse Transcription Kit (Thermo Fisher), with the associated RT primer pool, or High-Capacity cDNA Archive Kit (Thermo Fisher) were used to retrotranscribe miRs and mRNAs, respectively; TaqMan PreAmp Master Mix (Thermo Fisher) with the associated pre-amplification primer pool was used for pre-amplifications. miR-574-5p expression was assessed by Qiagen assays (Qiagen, Hilden, Germany), after retrotranscription by miScript II RT Kit (Thermo Fisher) and pre-amplification by miScript PreAMP PCR Kit (Thermo Fisher) using as internal reference U6 snRNA (Qiagen, Hilden, Germany). .. TNFRSF8 , TNF Receptor Superfamily Member 8 (Hs00174277_m1) and Beta Actin (4326315E) TaqMan Gene Expression assays were used.

    Article Title: Target Genes of Neuron-Restrictive Silencer Factor Are Abnormally Up-Regulated in Human Myotilinopathy
    Article Snippet: .. TaqMan PCR assays for α-internexin and SNAP25 from human muscle biopsies were performed using the TaqMan PreAmp Master Kit (Applied Biosystems). .. Chromatin immunoprecipitation (ChIP) assay was performed according to the manufacturer’s protocol (Upstate, Madrid, Spain) using 106 U87-MG, HeLa, and DMS53 cells.

    Article Title: A novel microRNA mmu-mir-466h affects apoptosis regulation in mammalian cells
    Article Snippet: .. The preamplification (10 cycles) was done using Applied Systems TaqMan® PreAmp Kit (Part No. 4384267) in PCR Thermal cycler (Applied Biosystems) after miRNA reverse transcription and before qRT-PCR reads. .. TaqMan® PreAmp Master Mix preamplifies small amounts of cDNA without introducing amplification bias to the sample and provides a very high correlation coefficients between amplified and unamplified cDNA ( ). qRT-PCR measurements were performed in quadruplicates.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher taqman master mix
    Inflammatory cytokine and Wnt mRNA levels in primary human 3D lung aggregate cultures and human macrophages (M) after cigarette smoke extracts (CSE) exposure. (A) IL-6 and IL-8 inflammatory cytokine mRNA levels in control and CSE exposed (48 h) lung aggregate cultures, containing and not containing M. (B) IL-6 and IL-8 inflammatory cytokine mRNA levels in control and rWnt5a (1 µg/ml) treated lung aggregate cultures, containing and not containing M. (C) <t>Taqman</t> array heat map analysis of pooled ( n = 4) <t>cDNA</t> of human M compared with control. M were treated with CSE for 3 h. (D) Changes in mRNA levels of Wnt signaling pathway genes measured by Taqman Array analysis using pooled ( n = 4) cDNA samples of human M compared with control.
    Taqman Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 692 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taqman master mix/product/Thermo Fisher
    Average 99 stars, based on 692 article reviews
    Price from $9.99 to $1999.99
    taqman master mix - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    94
    Thermo Fisher taqman preamp cells to ct kit
    Targeting the dominant ALDH isoform in high AVS HNSCC depletes the CIC pool. UMSCC47 and SCC25 cells were transduced with the inducible pLV-RNAi/shRNA-ALDH1A3 and polyclonal cell populations were collected. Cells were stimulated with doxycycline at 1000 ng/ml for all the experiments. ( a ) ALDH1A3 protein levels. Cell lysates were immunoblotted with anti-ALDH1A3 and GAPDH antibodies. Representative image is cropped. ( b ) ALDH1A3 <t>mRNA</t> expression. ALDH1A3 and GAPDH expression was determined using qPCR with <t>TaqMan</t> primers. Data were normalized to GAPDH and are presented as mean ± s.e.m. (n = 3, *p
    Taqman Preamp Cells To Ct Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taqman preamp cells to ct kit/product/Thermo Fisher
    Average 94 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    taqman preamp cells to ct kit - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    90
    Thermo Fisher miscript preamp pcr kit
    Targeting the dominant ALDH isoform in high AVS HNSCC depletes the CIC pool. UMSCC47 and SCC25 cells were transduced with the inducible pLV-RNAi/shRNA-ALDH1A3 and polyclonal cell populations were collected. Cells were stimulated with doxycycline at 1000 ng/ml for all the experiments. ( a ) ALDH1A3 protein levels. Cell lysates were immunoblotted with anti-ALDH1A3 and GAPDH antibodies. Representative image is cropped. ( b ) ALDH1A3 <t>mRNA</t> expression. ALDH1A3 and GAPDH expression was determined using qPCR with <t>TaqMan</t> primers. Data were normalized to GAPDH and are presented as mean ± s.e.m. (n = 3, *p
    Miscript Preamp Pcr Kit, 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
    https://www.bioz.com/result/miscript preamp pcr kit/product/Thermo Fisher
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    miscript preamp pcr kit - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    Image Search Results


    Inflammatory cytokine and Wnt mRNA levels in primary human 3D lung aggregate cultures and human macrophages (M) after cigarette smoke extracts (CSE) exposure. (A) IL-6 and IL-8 inflammatory cytokine mRNA levels in control and CSE exposed (48 h) lung aggregate cultures, containing and not containing M. (B) IL-6 and IL-8 inflammatory cytokine mRNA levels in control and rWnt5a (1 µg/ml) treated lung aggregate cultures, containing and not containing M. (C) Taqman array heat map analysis of pooled ( n = 4) cDNA of human M compared with control. M were treated with CSE for 3 h. (D) Changes in mRNA levels of Wnt signaling pathway genes measured by Taqman Array analysis using pooled ( n = 4) cDNA samples of human M compared with control.

    Journal: Frontiers in Immunology

    Article Title: Cigarette Smoke-Induced Pulmonary Inflammation Becomes Systemic by Circulating Extracellular Vesicles Containing Wnt5a and Inflammatory Cytokines

    doi: 10.3389/fimmu.2018.01724

    Figure Lengend Snippet: Inflammatory cytokine and Wnt mRNA levels in primary human 3D lung aggregate cultures and human macrophages (M) after cigarette smoke extracts (CSE) exposure. (A) IL-6 and IL-8 inflammatory cytokine mRNA levels in control and CSE exposed (48 h) lung aggregate cultures, containing and not containing M. (B) IL-6 and IL-8 inflammatory cytokine mRNA levels in control and rWnt5a (1 µg/ml) treated lung aggregate cultures, containing and not containing M. (C) Taqman array heat map analysis of pooled ( n = 4) cDNA of human M compared with control. M were treated with CSE for 3 h. (D) Changes in mRNA levels of Wnt signaling pathway genes measured by Taqman Array analysis using pooled ( n = 4) cDNA samples of human M compared with control.

    Article Snippet: TaqMan master mix was combined with the cDNA samples then the mixed solutions were loaded onto the Human WNT Pathway, Fast 96-well TaqMan Array (Thermo Fisher Scientific, Waltham, MA, USA) plate.

    Techniques:

    miTRAP as a strategy to purify Pax6 3’UTR-associated miRNAs. a Schematic of the Pax6 3’UTR affinity purification approach. (i) Plasmid vectors expressing GFP tagged with the MS2 RNA sequence motif followed by the SV40 polyadenylation signal are introduced into pancreatic αTC1–6 cells via transient transfection. (ii) MS2 coat protein fused to maltose binding (MS2-MBP) is used to purify GFP transcripts with bound miRNAs from αTC1–6 cell lysate. (iii) Real-time quantitative PCR (RT-qPCR) is used to detect GFP transcript and bound miRNAs. Schematic of the Pax6 3’UTR shows the location of the highly conserved miR-375 target site located at 3’UTR position 201 and miR-375 target site mutation. b ]. qPCR results for GFP with the MS2 motif (grey bar) were expressed relative to data without the MS2 motif (unfilled bar). Data represents 5 independent samples, p = 0.0079. c Affinity purification of miR-375 with the Pax6 3’UTR in αTC1–6 cells using TaqMan individual qPCR assays. Normalized relative quantity was calculated using Pfaffl’s method, and a normalized relative quantity greater than 1 indicates that more target miRNA is purified with the Pax6 3’UTR (grey bar) than the control lacking a Pax6 3’UTR (unfilled bar). Data represents six independent samples, p = 0.013. d Disruption of miR-375 binding to the Pax6 3’UTR following mutation of the miR-375 target site. Target miR-375 values were normalized to GFP as a reference gene, then normalized values for the mutant Pax6 3’UTR samples (grey bar), and are presented relative to the wt 3’UTR (unfilled bar). Data represents six independent wt 3’UTR samples and three miR-375 target site mutant 3’UTR samples, p = 0.0476. Error bars represent 95% confidence intervals, and p -values were calculated using the Mann Whitney test. Note scale bar differences between the graphs

    Journal: BMC Genomics

    Article Title: Mapping the Pax6 3’ untranslated region microRNA regulatory landscape

    doi: 10.1186/s12864-018-5212-x

    Figure Lengend Snippet: miTRAP as a strategy to purify Pax6 3’UTR-associated miRNAs. a Schematic of the Pax6 3’UTR affinity purification approach. (i) Plasmid vectors expressing GFP tagged with the MS2 RNA sequence motif followed by the SV40 polyadenylation signal are introduced into pancreatic αTC1–6 cells via transient transfection. (ii) MS2 coat protein fused to maltose binding (MS2-MBP) is used to purify GFP transcripts with bound miRNAs from αTC1–6 cell lysate. (iii) Real-time quantitative PCR (RT-qPCR) is used to detect GFP transcript and bound miRNAs. Schematic of the Pax6 3’UTR shows the location of the highly conserved miR-375 target site located at 3’UTR position 201 and miR-375 target site mutation. b ]. qPCR results for GFP with the MS2 motif (grey bar) were expressed relative to data without the MS2 motif (unfilled bar). Data represents 5 independent samples, p = 0.0079. c Affinity purification of miR-375 with the Pax6 3’UTR in αTC1–6 cells using TaqMan individual qPCR assays. Normalized relative quantity was calculated using Pfaffl’s method, and a normalized relative quantity greater than 1 indicates that more target miRNA is purified with the Pax6 3’UTR (grey bar) than the control lacking a Pax6 3’UTR (unfilled bar). Data represents six independent samples, p = 0.013. d Disruption of miR-375 binding to the Pax6 3’UTR following mutation of the miR-375 target site. Target miR-375 values were normalized to GFP as a reference gene, then normalized values for the mutant Pax6 3’UTR samples (grey bar), and are presented relative to the wt 3’UTR (unfilled bar). Data represents six independent wt 3’UTR samples and three miR-375 target site mutant 3’UTR samples, p = 0.0476. Error bars represent 95% confidence intervals, and p -values were calculated using the Mann Whitney test. Note scale bar differences between the graphs

    Article Snippet: miTRAP cDNA for use with TaqMan miRNA multiplex arrays was first preamplified using TaqMan PreAmp Master Mix (ThermoFisher, 4,391,128) and custom miRNA PreAmp primer pool.

    Techniques: Affinity Purification, Plasmid Preparation, Expressing, Sequencing, Transfection, Binding Assay, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Mutagenesis, Purification, MANN-WHITNEY

    Targeting the dominant ALDH isoform in high AVS HNSCC depletes the CIC pool. UMSCC47 and SCC25 cells were transduced with the inducible pLV-RNAi/shRNA-ALDH1A3 and polyclonal cell populations were collected. Cells were stimulated with doxycycline at 1000 ng/ml for all the experiments. ( a ) ALDH1A3 protein levels. Cell lysates were immunoblotted with anti-ALDH1A3 and GAPDH antibodies. Representative image is cropped. ( b ) ALDH1A3 mRNA expression. ALDH1A3 and GAPDH expression was determined using qPCR with TaqMan primers. Data were normalized to GAPDH and are presented as mean ± s.e.m. (n = 3, *p

    Journal: Scientific Reports

    Article Title: p53 functional states are associated with distinct aldehyde dehydrogenase transcriptomic signatures

    doi: 10.1038/s41598-020-57758-5

    Figure Lengend Snippet: Targeting the dominant ALDH isoform in high AVS HNSCC depletes the CIC pool. UMSCC47 and SCC25 cells were transduced with the inducible pLV-RNAi/shRNA-ALDH1A3 and polyclonal cell populations were collected. Cells were stimulated with doxycycline at 1000 ng/ml for all the experiments. ( a ) ALDH1A3 protein levels. Cell lysates were immunoblotted with anti-ALDH1A3 and GAPDH antibodies. Representative image is cropped. ( b ) ALDH1A3 mRNA expression. ALDH1A3 and GAPDH expression was determined using qPCR with TaqMan primers. Data were normalized to GAPDH and are presented as mean ± s.e.m. (n = 3, *p

    Article Snippet: Quantitative real-time PCR Cells were extracted for total RNA using the TRIzol reagent (ThermoFisher Scientific, Waltham, MA) or TaqMan PreAmp Cells-to-CT kit (ThermoFisher Scientific). mRNA expression of ALDH isoforms was determined using the Applied Biosystems 7900HT Fast Real-Time PCR System with validated, pre-designed TaqMan primers (ThermoFisher Scientific).

    Techniques: Transduction, shRNA, Expressing, Real-time Polymerase Chain Reaction