superscript iii kit  (Thermo Fisher)


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    Structured Review

    Thermo Fisher superscript iii kit
    Cloning strategy used for Scdr2 overexpression in tobacco plants. (A) The complete coding sequence of Scdr2 was cloned under the control of the constitutive Cauliflower mosaic virus (CaMV) 35S promoter (p35S) and with the NOS polyadenylation signal (Nos-t) using pCambia2301 as the backbone. nptII (kanamycin resistance) gene expression was also driven by the p35S promoter. LB and RB correspond to the T-DNA left and right borders, respectively. Positions of some of the restriction sites are indicated. (B) Expression of Scdr2 in <t>three</t> independent T 3 generation transgenic lines and the WT. Total <t>RNA</t> was extracted from 2-week-old seedlings and then analysed using semi-quantitative RT–PCR. The Scdr2 gene product was obtained after 21 cycles, while the product of the tobacco actin gene, which was used as an internal standard, was obtained after 28 cycles ( n = 3). (C) Densitometric analysis of Scdr2 expression that was obtained from the semi-quantitative RT–PCR shown in (B).
    Superscript Iii Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii kit/product/Thermo Fisher
    Average 99 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    superscript iii kit - by Bioz Stars, 2020-01
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    Images

    1) Product Images from "Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience"

    Article Title: Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience

    Journal: Annals of Botany

    doi: 10.1093/aob/mcz044

    Cloning strategy used for Scdr2 overexpression in tobacco plants. (A) The complete coding sequence of Scdr2 was cloned under the control of the constitutive Cauliflower mosaic virus (CaMV) 35S promoter (p35S) and with the NOS polyadenylation signal (Nos-t) using pCambia2301 as the backbone. nptII (kanamycin resistance) gene expression was also driven by the p35S promoter. LB and RB correspond to the T-DNA left and right borders, respectively. Positions of some of the restriction sites are indicated. (B) Expression of Scdr2 in three independent T 3 generation transgenic lines and the WT. Total RNA was extracted from 2-week-old seedlings and then analysed using semi-quantitative RT–PCR. The Scdr2 gene product was obtained after 21 cycles, while the product of the tobacco actin gene, which was used as an internal standard, was obtained after 28 cycles ( n = 3). (C) Densitometric analysis of Scdr2 expression that was obtained from the semi-quantitative RT–PCR shown in (B).
    Figure Legend Snippet: Cloning strategy used for Scdr2 overexpression in tobacco plants. (A) The complete coding sequence of Scdr2 was cloned under the control of the constitutive Cauliflower mosaic virus (CaMV) 35S promoter (p35S) and with the NOS polyadenylation signal (Nos-t) using pCambia2301 as the backbone. nptII (kanamycin resistance) gene expression was also driven by the p35S promoter. LB and RB correspond to the T-DNA left and right borders, respectively. Positions of some of the restriction sites are indicated. (B) Expression of Scdr2 in three independent T 3 generation transgenic lines and the WT. Total RNA was extracted from 2-week-old seedlings and then analysed using semi-quantitative RT–PCR. The Scdr2 gene product was obtained after 21 cycles, while the product of the tobacco actin gene, which was used as an internal standard, was obtained after 28 cycles ( n = 3). (C) Densitometric analysis of Scdr2 expression that was obtained from the semi-quantitative RT–PCR shown in (B).

    Techniques Used: Clone Assay, Over Expression, Sequencing, Expressing, Transgenic Assay, Quantitative RT-PCR

    2) Product Images from "Rat NaV1.7 loss-of-function genetic model: Deficient nociceptive and neuropathic pain behavior with retained olfactory function and intra-epidermal nerve fibers"

    Article Title: Rat NaV1.7 loss-of-function genetic model: Deficient nociceptive and neuropathic pain behavior with retained olfactory function and intra-epidermal nerve fibers

    Journal: Molecular Pain

    doi: 10.1177/1744806919881846

    Na V 1.7 Immunostaining in DRG and sciatic nerve. (A and A′) Na V 1.7 immunoreactivity in DRG from WT but not HOM-KI rats. (B and B′). Corresponding isotype control sections. (C and C′) Na V 1.7 immunoreactivity in sciatic nerve from WT but not HOM-KI rats. (D and D′) Corresponding isotype control sections. Scale bars 200 µm. Three rats of each genotype and gender were evaluated. HOM-KI: rats homozygous for the knock-in allele; WT: wild type.
    Figure Legend Snippet: Na V 1.7 Immunostaining in DRG and sciatic nerve. (A and A′) Na V 1.7 immunoreactivity in DRG from WT but not HOM-KI rats. (B and B′). Corresponding isotype control sections. (C and C′) Na V 1.7 immunoreactivity in sciatic nerve from WT but not HOM-KI rats. (D and D′) Corresponding isotype control sections. Scale bars 200 µm. Three rats of each genotype and gender were evaluated. HOM-KI: rats homozygous for the knock-in allele; WT: wild type.

    Techniques Used: Immunostaining, Knock-In

    3) Product Images from "A novel small-form NEDD4 regulates cell invasiveness and apoptosis to promote tumor metastasis"

    Article Title: A novel small-form NEDD4 regulates cell invasiveness and apoptosis to promote tumor metastasis

    Journal: Oncotarget

    doi:

    Generation of potent metastatic cell lines, SKT and SKM, and identifying metastatic-associated gene, sNEDD4 (A) Schematic diagram of the experimental procedure. (B) The invasive abilities of SK, SKT, and SKM cells were determined with the Transwell invasion assay. (C) Western blot analysis was used to determined NEDD4 protein expression of SK, SKT, and SKM cells. Three (left; upper, middle, and lower) and two (right; upper and lower) major bands were detected using NEDD4 and NEDD4–1 antibodies in SKM cells, respectively. (D) Schematic diagram of NEDD4 mRNA (Upper). The open box represents the coding region of NEDD4 . Classic and predictive ATG sites are labeled with gray bars. The lower schematic diagram represents NEDD4 and sNEDD4 protein structures. (E) NEDD4 mRNA levels were determined via qRT-PCR with primers amplified from two different segments, S1 and S2 (mark in panel D). (F) NEDD4 and sNEDD4 protein levels of SK, SKM and SK cells transiently transfected with empty vector (Vec) or sNEDD4 expression plasmid (sNED). (G) sNEDD4 was detected using Northern blot. SK cells transfected with sNEDD4 expressing plasmid (sNED) serve as a positive control. IB, immunobloting; S1, 366–478 nucleotides; S2, 649–751 nucleotides; *** P
    Figure Legend Snippet: Generation of potent metastatic cell lines, SKT and SKM, and identifying metastatic-associated gene, sNEDD4 (A) Schematic diagram of the experimental procedure. (B) The invasive abilities of SK, SKT, and SKM cells were determined with the Transwell invasion assay. (C) Western blot analysis was used to determined NEDD4 protein expression of SK, SKT, and SKM cells. Three (left; upper, middle, and lower) and two (right; upper and lower) major bands were detected using NEDD4 and NEDD4–1 antibodies in SKM cells, respectively. (D) Schematic diagram of NEDD4 mRNA (Upper). The open box represents the coding region of NEDD4 . Classic and predictive ATG sites are labeled with gray bars. The lower schematic diagram represents NEDD4 and sNEDD4 protein structures. (E) NEDD4 mRNA levels were determined via qRT-PCR with primers amplified from two different segments, S1 and S2 (mark in panel D). (F) NEDD4 and sNEDD4 protein levels of SK, SKM and SK cells transiently transfected with empty vector (Vec) or sNEDD4 expression plasmid (sNED). (G) sNEDD4 was detected using Northern blot. SK cells transfected with sNEDD4 expressing plasmid (sNED) serve as a positive control. IB, immunobloting; S1, 366–478 nucleotides; S2, 649–751 nucleotides; *** P

    Techniques Used: Transwell Invasion Assay, Western Blot, Expressing, Labeling, Quantitative RT-PCR, Amplification, Transfection, Plasmid Preparation, Northern Blot, Positive Control

    Related Articles

    SYBR Green Assay:

    Article Title: Inflammasome activation in airway epithelial cells after multi-walled carbon nanotube exposure mediates a profibrotic response in lung fibroblasts
    Article Snippet: Reverse transcription (RT) reaction to obtain cDNA was performed with SuperScript III kit and oligo dTs (Invitrogen) exactly following manufacturer’s recommendations. .. Final PCR reaction volume was 25 μL containing Power SYBR Green Master Mix (Applied Biosciences), 1 μg of cDNA and 500 nM of each specific forward and reverse primer (Table ).

    Article Title: A novel small-form NEDD4 regulates cell invasiveness and apoptosis to promote tumor metastasis
    Article Snippet: qRT-PCR Total RNA was extracted and cDNA strands synthesized using the Superscript III kit for RT-PCR (Life Technologies, USA). .. Real-time quantitative RT-PCR was performed using SYBR green, and detected using the ABI PRISM 7500 system (Applied Biosystems, UK).

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific). .. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was carried out with QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific) using Power SYBR Green PCR Master Mix (Cat. 4367659, Thermo Fisher Scientific Inc, USA).

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: .. RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents. ..

    Article Title: GRAINYHEAD-LIKE-2 CONFERS NK-SENSITIVITY THROUGH INTERACTIONS WITH EPIGENETIC MODIFIERS
    Article Snippet: RNA (2.5 μg) were converted to single strand cDNA using the Superscript III kit (Life Technologies). .. One microliter of (3:1 diluted) cDNA was assayed in 20 uL reactions using the PowerUp SYBR green master mix (Applied Biosystems) on an Applied Biosystems 7500 thermocycler.

    Cell Isolation:

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: RNA from CD4-enriched T cells (CD4 T Cell Isolation Kit, Miltenyi Biotech) was isolated using TRIzol Reagent (Cat. 15596–026, Thermo Fisher Scientific). .. First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific).

    Amplification:

    Article Title: Identification of a Novel Betacoronavirus (Merbecovirus) in Amur Hedgehogs from China
    Article Snippet: Reverse transcription was carried out according to the suggested protocol of the SuperScript III kit (Invitrogen, Life Technologies, Grand Island, NY, USA). .. The amplification of 25 μL PCR mixtures was carried out in automated thermal cyclers (Applied Biosystems).

    Article Title: The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1 [W]The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1 [W] [OA]
    Article Snippet: .. Following RNA isolation, complementary DNA ( ) was made from the total RNA using the SuperScript III kit for -PCR from Invitrogen according to the manufacturer’s recommendations. was performed with SuperScript III reverse transcriptase followed by PCR amplification of the . .. Quantitative real-time PCR was carried out using 2X QuantiTect SYBR Green PCR Master Mix on an Applied Biosystems Fast System 7500 cycler.

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: Primers used for preamplification and quantitative measurement were intron-spanning and validated on serial dilution of total mouse cDNA to ensure linear amplification and PCR specificity. .. RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents.

    Expressing:

    Article Title: Inflammasome activation in airway epithelial cells after multi-walled carbon nanotube exposure mediates a profibrotic response in lung fibroblasts
    Article Snippet: Reverse transcription (RT) reaction to obtain cDNA was performed with SuperScript III kit and oligo dTs (Invitrogen) exactly following manufacturer’s recommendations. .. Changes in expression of target genes were determined by ddCt analysis, comparing expression to Ribosomal Protein (18 s) and normalizing to media control.

    Article Title: Autosomal recessive congenital cataracts linked to HSF4 in a consanguineous Pakistani family
    Article Snippet: Paragraph title: Real-time expression analysis ... First-strand cDNA synthesis was completed using the Superscript III kit (Invitrogen) according to the manufacturer’s instructions.

    Article Title: A Functional Variant at a Prostate Cancer Predisposition Locus at 8q24 Is Associated with PVT1 Expression
    Article Snippet: .. mRNA expression analysis cDNA was prepared from 800 ng of total RNA with Superscript III kit and random hexamers (Invitrogen). .. All expression assays were first evaluated in pooled prostate cDNA samples containing 25 ng, 5 ng, or 1 ng of total RNA per reaction.

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific). .. As an internal control, hypoxanthine guanine phosphoribosyl transferase (HPRT) RNA levels were used to normalize gene expression.

    Article Title: Modulation of GSK-3 provides cellular and functional neuroprotection in the rd10 mouse model of retinitis pigmentosa
    Article Snippet: Total RNA from individual retinas was extracted using TRIzol Reagent, and 2.5 μg of RNA was typically reverse transcribed using the Superscript III Kit and random primers (all from ThermoFisher Scientific). .. The relative change in gene expression was calculated using the 2ΔCt method, normalizing to expression levels of the Tbp (TATA-binding protein) gene.

    Article Title: SUMO3 modification by PIAS1 modulates androgen receptor cellular distribution and stability
    Article Snippet: Two μg total RNA were transcribed (RT) into 20 μl cDNA by SuperScript III kit (Invitrogen) with oligo (dT) primer. .. We designed AR primers for PCR to measure its gene expression level and used the β-actin expression level as control.

    Article Title: Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience
    Article Snippet: First-strand cDNA was produced from 2 μg of total RNA using oligo d(T)18 primers and the Superscript III Kit according to the manufacturer’s instructions (Life Technologies, USA). .. The 2–ΔΔCt method ( ) was used to calculate the relative expression of genes (experimental/control) using a sample that was not subjected to stress as a control and a gene encoding polyubiquitin as a reference.

    Synthesized:

    Article Title: A novel small-form NEDD4 regulates cell invasiveness and apoptosis to promote tumor metastasis
    Article Snippet: .. qRT-PCR Total RNA was extracted and cDNA strands synthesized using the Superscript III kit for RT-PCR (Life Technologies, USA). .. Real-time quantitative RT-PCR was performed using SYBR green, and detected using the ABI PRISM 7500 system (Applied Biosystems, UK).

    Article Title: PTBP1-mediated regulation of AXL mRNA stability plays a role in lung tumorigenesis
    Article Snippet: .. The first strand of cDNA was synthesized using the Superscript III kit (Invitrogen). .. PCR reactions were performed in a 20 μl reaction mixture containing 10 μM forward and reverse primers, 1X SYBER GREEN reaction mix (Takara).

    Isolation:

    Article Title: Autosomal recessive congenital cataracts linked to HSF4 in a consanguineous Pakistani family
    Article Snippet: Lenses were dissolved in Trizol reagent (Invitrogen; Carlsbad, CA) immediately after extraction and total RNA was isolated according to the manufacturer’s instructions. .. First-strand cDNA synthesis was completed using the Superscript III kit (Invitrogen) according to the manufacturer’s instructions.

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: Paragraph title: RNA isolation, reverse transcription and quantitative polymerase chain reaction (qPCR). ... First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific).

    Article Title: The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1 [W]The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1 [W] [OA]
    Article Snippet: .. Following RNA isolation, complementary DNA ( ) was made from the total RNA using the SuperScript III kit for -PCR from Invitrogen according to the manufacturer’s recommendations. was performed with SuperScript III reverse transcriptase followed by PCR amplification of the . .. Quantitative real-time PCR was carried out using 2X QuantiTect SYBR Green PCR Master Mix on an Applied Biosystems Fast System 7500 cycler.

    Article Title: p75NTR antagonists attenuate photoreceptor cell loss in murine models of retinitis pigmentosa
    Article Snippet: .. RNA isolation and quantitative PCR Total RNA from individual retinas was extracted using the TRIzol reagent, and 2.5 μ g of RNA were typically reverse transcribed (RT) using the Superscript III Kit and random primers (all from Thermo Fisher Scientific). .. Quantitative PCR (qPCR) was performed with the ABI Prism 7900HT Sequence Detection System using TaqMan Universal PCR Master Mix, No-AmpErase UNG and the Taqman assays (listed below) for detection (all from Thermo Fisher).

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: Results were validated by classical qRT-PCR using total RNA isolated from ∼0.5–2 × 104 cells. .. RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents.

    Article Title: Modulation of GSK-3 provides cellular and functional neuroprotection in the rd10 mouse model of retinitis pigmentosa
    Article Snippet: Paragraph title: RNA isolation and quantitative PCR ... Total RNA from individual retinas was extracted using TRIzol Reagent, and 2.5 μg of RNA was typically reverse transcribed using the Superscript III Kit and random primers (all from ThermoFisher Scientific).

    Negative Control:

    Article Title: Rat NaV1.7 loss-of-function genetic model: Deficient nociceptive and neuropathic pain behavior with retained olfactory function and intra-epidermal nerve fibers
    Article Snippet: cDNA synthesis Furthermore, 500 ng of total RNA was taken from each DRG sample for synthesis of first-strand cDNA using the SuperScript™ III kit (Invitrogen Cat no. 18080–093). .. SuperScript reverse transcriptase was replaced by an equal volume of nuclease-free water for the generation of a reverse transcriptase negative control for each sample.

    Microscopy:

    Article Title: Autosomal recessive congenital cataracts linked to HSF4 in a consanguineous Pakistani family
    Article Snippet: The ocular tissue was extracted, and the lenses were isolated from retina using forceps under a microscope. .. First-strand cDNA synthesis was completed using the Superscript III kit (Invitrogen) according to the manufacturer’s instructions.

    Mouse Assay:

    Article Title: Autosomal recessive congenital cataracts linked to HSF4 in a consanguineous Pakistani family
    Article Snippet: Mice were first anesthetized by isoflurane and subsequently euthanized through cervical dislocation. .. First-strand cDNA synthesis was completed using the Superscript III kit (Invitrogen) according to the manufacturer’s instructions.

    Real-time Polymerase Chain Reaction:

    Article Title: Inflammasome activation in airway epithelial cells after multi-walled carbon nanotube exposure mediates a profibrotic response in lung fibroblasts
    Article Snippet: Reverse transcription (RT) reaction to obtain cDNA was performed with SuperScript III kit and oligo dTs (Invitrogen) exactly following manufacturer’s recommendations. .. Data was collected on a Stratagene Mx3005P real-time PCR instrument (Agilent, CA, USA).

    Article Title: Autosomal recessive congenital cataracts linked to HSF4 in a consanguineous Pakistani family
    Article Snippet: First-strand cDNA synthesis was completed using the Superscript III kit (Invitrogen) according to the manufacturer’s instructions. .. Quantitative real-time PCR analyses were performed on STEP ONE ABI Real-Time PCR System using predesigned Hsf4 TaqMan expression assays (Applied Biosystems).

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: Paragraph title: RNA isolation, reverse transcription and quantitative polymerase chain reaction (qPCR). ... First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific).

    Article Title: p75NTR antagonists attenuate photoreceptor cell loss in murine models of retinitis pigmentosa
    Article Snippet: .. RNA isolation and quantitative PCR Total RNA from individual retinas was extracted using the TRIzol reagent, and 2.5 μ g of RNA were typically reverse transcribed (RT) using the Superscript III Kit and random primers (all from Thermo Fisher Scientific). .. Quantitative PCR (qPCR) was performed with the ABI Prism 7900HT Sequence Detection System using TaqMan Universal PCR Master Mix, No-AmpErase UNG and the Taqman assays (listed below) for detection (all from Thermo Fisher).

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: .. RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents. ..

    Article Title: Modulation of GSK-3 provides cellular and functional neuroprotection in the rd10 mouse model of retinitis pigmentosa
    Article Snippet: Paragraph title: RNA isolation and quantitative PCR ... Total RNA from individual retinas was extracted using TRIzol Reagent, and 2.5 μg of RNA was typically reverse transcribed using the Superscript III Kit and random primers (all from ThermoFisher Scientific).

    Article Title: Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience
    Article Snippet: Paragraph title: Real-time qPCR and semi-quantitative RT–PCR ... First-strand cDNA was produced from 2 μg of total RNA using oligo d(T)18 primers and the Superscript III Kit according to the manufacturer’s instructions (Life Technologies, USA).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Identification of a Novel Betacoronavirus (Merbecovirus) in Amur Hedgehogs from China
    Article Snippet: Paragraph title: 2.3. Detection of CoVs by RT-PCR and Sequencing ... Reverse transcription was carried out according to the suggested protocol of the SuperScript III kit (Invitrogen, Life Technologies, Grand Island, NY, USA).

    Article Title: A novel small-form NEDD4 regulates cell invasiveness and apoptosis to promote tumor metastasis
    Article Snippet: .. qRT-PCR Total RNA was extracted and cDNA strands synthesized using the Superscript III kit for RT-PCR (Life Technologies, USA). .. Real-time quantitative RT-PCR was performed using SYBR green, and detected using the ABI PRISM 7500 system (Applied Biosystems, UK).

    Article Title: SUMO3 modification by PIAS1 modulates androgen receptor cellular distribution and stability
    Article Snippet: Paragraph title: RNA extraction and RT-PCR ... Two μg total RNA were transcribed (RT) into 20 μl cDNA by SuperScript III kit (Invitrogen) with oligo (dT) primer.

    Article Title: Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience
    Article Snippet: Paragraph title: Real-time qPCR and semi-quantitative RT–PCR ... First-strand cDNA was produced from 2 μg of total RNA using oligo d(T)18 primers and the Superscript III Kit according to the manufacturer’s instructions (Life Technologies, USA).

    Polymerase Chain Reaction:

    Article Title: Inflammasome activation in airway epithelial cells after multi-walled carbon nanotube exposure mediates a profibrotic response in lung fibroblasts
    Article Snippet: Reverse transcription (RT) reaction to obtain cDNA was performed with SuperScript III kit and oligo dTs (Invitrogen) exactly following manufacturer’s recommendations. .. Final PCR reaction volume was 25 μL containing Power SYBR Green Master Mix (Applied Biosciences), 1 μg of cDNA and 500 nM of each specific forward and reverse primer (Table ).

    Article Title: Identification of a Novel Betacoronavirus (Merbecovirus) in Amur Hedgehogs from China
    Article Snippet: Reverse transcription was carried out according to the suggested protocol of the SuperScript III kit (Invitrogen, Life Technologies, Grand Island, NY, USA). .. The amplification of 25 μL PCR mixtures was carried out in automated thermal cyclers (Applied Biosystems).

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific). .. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was carried out with QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific) using Power SYBR Green PCR Master Mix (Cat. 4367659, Thermo Fisher Scientific Inc, USA).

    Article Title: The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1 [W]The Role of TIR-NBS and TIR-X Proteins in Plant Basal Defense Responses 1 [W] [OA]
    Article Snippet: .. Following RNA isolation, complementary DNA ( ) was made from the total RNA using the SuperScript III kit for -PCR from Invitrogen according to the manufacturer’s recommendations. was performed with SuperScript III reverse transcriptase followed by PCR amplification of the . .. Quantitative real-time PCR was carried out using 2X QuantiTect SYBR Green PCR Master Mix on an Applied Biosystems Fast System 7500 cycler.

    Article Title: p75NTR antagonists attenuate photoreceptor cell loss in murine models of retinitis pigmentosa
    Article Snippet: RNA isolation and quantitative PCR Total RNA from individual retinas was extracted using the TRIzol reagent, and 2.5 μ g of RNA were typically reverse transcribed (RT) using the Superscript III Kit and random primers (all from Thermo Fisher Scientific). .. Quantitative PCR (qPCR) was performed with the ABI Prism 7900HT Sequence Detection System using TaqMan Universal PCR Master Mix, No-AmpErase UNG and the Taqman assays (listed below) for detection (all from Thermo Fisher).

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: Primers used for preamplification and quantitative measurement were intron-spanning and validated on serial dilution of total mouse cDNA to ensure linear amplification and PCR specificity. .. RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents.

    Article Title: Modulation of GSK-3 provides cellular and functional neuroprotection in the rd10 mouse model of retinitis pigmentosa
    Article Snippet: Total RNA from individual retinas was extracted using TRIzol Reagent, and 2.5 μg of RNA was typically reverse transcribed using the Superscript III Kit and random primers (all from ThermoFisher Scientific). .. Quantitative PCR (qPCR) was performed with the ABI Prism 7900HT Sequence Detection System using TaqMan Universal PCR Master Mix, no-AmpErase UNG, and Taqman assays (listed below) for detection (all from Thermo Fisher).

    Article Title: SUMO3 modification by PIAS1 modulates androgen receptor cellular distribution and stability
    Article Snippet: RNA extraction and RT-PCR RNA extraction and reverser transcriptional PCR analysis were used to quantify the change of AR mRNA levels induced by PIAS1 and SUMO3 at different time points. .. Two μg total RNA were transcribed (RT) into 20 μl cDNA by SuperScript III kit (Invitrogen) with oligo (dT) primer.

    Article Title: PTBP1-mediated regulation of AXL mRNA stability plays a role in lung tumorigenesis
    Article Snippet: The first strand of cDNA was synthesized using the Superscript III kit (Invitrogen). .. PCR reactions were performed in a 20 μl reaction mixture containing 10 μM forward and reverse primers, 1X SYBER GREEN reaction mix (Takara).

    Cell Culture:

    Article Title: SUMO3 modification by PIAS1 modulates androgen receptor cellular distribution and stability
    Article Snippet: We harvested cultured cells in TRIzol (Invitrogen) and extracted total RNA following the manufacturer’s instructions. .. Two μg total RNA were transcribed (RT) into 20 μl cDNA by SuperScript III kit (Invitrogen) with oligo (dT) primer.

    Quantitative RT-PCR:

    Article Title: Inflammasome activation in airway epithelial cells after multi-walled carbon nanotube exposure mediates a profibrotic response in lung fibroblasts
    Article Snippet: Paragraph title: Real time RT-PCR ... Reverse transcription (RT) reaction to obtain cDNA was performed with SuperScript III kit and oligo dTs (Invitrogen) exactly following manufacturer’s recommendations.

    Article Title: A novel small-form NEDD4 regulates cell invasiveness and apoptosis to promote tumor metastasis
    Article Snippet: .. qRT-PCR Total RNA was extracted and cDNA strands synthesized using the Superscript III kit for RT-PCR (Life Technologies, USA). .. Real-time quantitative RT-PCR was performed using SYBR green, and detected using the ABI PRISM 7500 system (Applied Biosystems, UK).

    Article Title: STAT6 and Furin Are Successive Triggers for the Production of TGFβ by T Cells
    Article Snippet: First-strand synthesis of cDNA from RNA was performed using SuperScript III kit (Cat. 18080–051, Thermo Fisher Scientific). .. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) was carried out with QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific) using Power SYBR Green PCR Master Mix (Cat. 4367659, Thermo Fisher Scientific Inc, USA).

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: Paragraph title: qRT-PCR array ... RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents.

    Article Title: PTBP1-mediated regulation of AXL mRNA stability plays a role in lung tumorigenesis
    Article Snippet: Paragraph title: Quantitative real-time RT-PCR ... The first strand of cDNA was synthesized using the Superscript III kit (Invitrogen).

    Article Title: GRAINYHEAD-LIKE-2 CONFERS NK-SENSITIVITY THROUGH INTERACTIONS WITH EPIGENETIC MODIFIERS
    Article Snippet: Paragraph title: qRT-PCR ... RNA (2.5 μg) were converted to single strand cDNA using the Superscript III kit (Life Technologies).

    Serial Dilution:

    Article Title: Obesity alters the long-term fitness of the hematopoietic stem cell compartment through modulation of Gfi1 expression
    Article Snippet: Primers used for preamplification and quantitative measurement were intron-spanning and validated on serial dilution of total mouse cDNA to ensure linear amplification and PCR specificity. .. RNA was treated with DNase I and reverse-transcribed using SuperScript III kit and random hexamers (Invitrogen). cDNA equivalent of 200 cells per reaction were used for quantitative PCR assays with SYBR green reagents.

    Sequencing:

    Article Title: Identification of a Novel Betacoronavirus (Merbecovirus) in Amur Hedgehogs from China
    Article Snippet: Paragraph title: 2.3. Detection of CoVs by RT-PCR and Sequencing ... Reverse transcription was carried out according to the suggested protocol of the SuperScript III kit (Invitrogen, Life Technologies, Grand Island, NY, USA).

    Article Title: p75NTR antagonists attenuate photoreceptor cell loss in murine models of retinitis pigmentosa
    Article Snippet: RNA isolation and quantitative PCR Total RNA from individual retinas was extracted using the TRIzol reagent, and 2.5 μ g of RNA were typically reverse transcribed (RT) using the Superscript III Kit and random primers (all from Thermo Fisher Scientific). .. Quantitative PCR (qPCR) was performed with the ABI Prism 7900HT Sequence Detection System using TaqMan Universal PCR Master Mix, No-AmpErase UNG and the Taqman assays (listed below) for detection (all from Thermo Fisher).

    Article Title: Modulation of GSK-3 provides cellular and functional neuroprotection in the rd10 mouse model of retinitis pigmentosa
    Article Snippet: Total RNA from individual retinas was extracted using TRIzol Reagent, and 2.5 μg of RNA was typically reverse transcribed using the Superscript III Kit and random primers (all from ThermoFisher Scientific). .. Quantitative PCR (qPCR) was performed with the ABI Prism 7900HT Sequence Detection System using TaqMan Universal PCR Master Mix, no-AmpErase UNG, and Taqman assays (listed below) for detection (all from Thermo Fisher).

    Article Title: PTBP1-mediated regulation of AXL mRNA stability plays a role in lung tumorigenesis
    Article Snippet: The first strand of cDNA was synthesized using the Superscript III kit (Invitrogen). .. All reactions were performed in an ABI 7500 sequence detection system.

    Produced:

    Article Title: Overexpression of an evolutionarily conserved drought-responsive sugarcane gene enhances salinity and drought resilience
    Article Snippet: .. First-strand cDNA was produced from 2 μg of total RNA using oligo d(T)18 primers and the Superscript III Kit according to the manufacturer’s instructions (Life Technologies, USA). .. The 2–ΔΔCt method ( ) was used to calculate the relative expression of genes (experimental/control) using a sample that was not subjected to stress as a control and a gene encoding polyubiquitin as a reference.

    Spectrophotometry:

    Article Title: Autosomal recessive congenital cataracts linked to HSF4 in a consanguineous Pakistani family
    Article Snippet: The quality and quantity of RNA were determined on a NanoDrop Lite spectrophotometer (Thermo Scientific, Inc.). .. First-strand cDNA synthesis was completed using the Superscript III kit (Invitrogen) according to the manufacturer’s instructions.

    RNA Extraction:

    Article Title: SUMO3 modification by PIAS1 modulates androgen receptor cellular distribution and stability
    Article Snippet: Paragraph title: RNA extraction and RT-PCR ... Two μg total RNA were transcribed (RT) into 20 μl cDNA by SuperScript III kit (Invitrogen) with oligo (dT) primer.

    Concentration Assay:

    Article Title: Rat NaV1.7 loss-of-function genetic model: Deficient nociceptive and neuropathic pain behavior with retained olfactory function and intra-epidermal nerve fibers
    Article Snippet: cDNA synthesis Furthermore, 500 ng of total RNA was taken from each DRG sample for synthesis of first-strand cDNA using the SuperScript™ III kit (Invitrogen Cat no. 18080–093). .. Upon completion of the synthesis reaction, each cDNA sample was diluted with 180 µl of IDTE pH 8.0 1× Tris-EDTA (TE) Solution (IDT Cat no. 11–05-01–13) to obtain a final concentration of 5 ng/µl.

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