superscript iii reverse transcriptase  (Millipore)


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    Millipore superscript iii reverse transcriptase
    Quantitative <t>PCR</t> validation of 12 DEGs in A375 cells treated with SFN. Relative fold expression of candidate genes was normalized against GADPH expression. The data represent the mean ± SD of <t>three</t> independent replicates. The black bars represent untreated cells at time 0, and the grey bars represent the relative fold change after SFN treatment. Statistical analysis was performed by paired two-tailed Student’s test (* p
    Superscript Iii Reverse Transcriptase, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript iii reverse transcriptase/product/Millipore
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    superscript iii reverse transcriptase - by Bioz Stars, 2020-04
    95/100 stars

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    1) Product Images from "Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells"

    Article Title: Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells

    Journal: European Journal of Nutrition

    doi: 10.1007/s00394-017-1527-7

    Quantitative PCR validation of 12 DEGs in A375 cells treated with SFN. Relative fold expression of candidate genes was normalized against GADPH expression. The data represent the mean ± SD of three independent replicates. The black bars represent untreated cells at time 0, and the grey bars represent the relative fold change after SFN treatment. Statistical analysis was performed by paired two-tailed Student’s test (* p
    Figure Legend Snippet: Quantitative PCR validation of 12 DEGs in A375 cells treated with SFN. Relative fold expression of candidate genes was normalized against GADPH expression. The data represent the mean ± SD of three independent replicates. The black bars represent untreated cells at time 0, and the grey bars represent the relative fold change after SFN treatment. Statistical analysis was performed by paired two-tailed Student’s test (* p

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Two Tailed Test

    Related Articles

    Clone Assay:

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: Paragraph title: Cloning of the DLac sequence ... Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer.

    Amplification:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. For non-quantitative expression analysis, cDNA was amplified by PCR for 35 cycles at 60°C as annealing temperature and amplified products were visualized after electrophoresis in a 1–2% agarose gel.

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer. .. To clone the laccase gene from the Cerrena sp. RSD1, the cDNA of DLac was specifically amplified using the degenerated primer NP2.

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates. ..

    Synthesized:

    Article Title: Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells
    Article Snippet: .. Quantitative real-time PCR (qPCR) cDNA was synthesized using Superscript III Reverse Transcriptase (Sigma) according to the manufacturer’s instructions. qPCR was performed in 96-well plates using iQ™ SYBR Green Supermix (Bio-Rad) and an I-cycler iQ Real-Time PCR instrument (Bio-Rad). .. Measurements were performed in triplicate with a variability < 0.5 Ct. mRNA expression was analysed by normalizing to that of the housekeeping gene GADPH.

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: .. Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: .. RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. All signals were quantified using the ACt method and were normalized to the levels of Gapdh.

    Cytometry:

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. For mCherry-positive flow cytometer sorted tumour and lung metastatic cells, cDNA was produced directly from lysed cells using a TaqMan Gene Expression Cells-to-Ct Kit (Thermo Fisher Scientific). qPCR was performed on an CFX96 (Bio-Rad) using TaqMan primer/probe sets and all signals were quantified as described above.

    Quantitative RT-PCR:

    Article Title: A model of breast cancer heterogeneity reveals vascular mimicry as a driver of metastasis
    Article Snippet: Paragraph title: qRT–PCR ... Synthesis of cDNA was performed using SuperScript III Reverse Transcriptase (Sigma).

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: .. Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. For non-quantitative expression analysis, cDNA was amplified by PCR for 35 cycles at 60°C as annealing temperature and amplified products were visualized after electrophoresis in a 1–2% agarose gel.

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: Paragraph title: Quantitative RT-PCR ... RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex.

    Electrophoresis:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. For non-quantitative expression analysis, cDNA was amplified by PCR for 35 cycles at 60°C as annealing temperature and amplified products were visualized after electrophoresis in a 1–2% agarose gel.

    Random Hexamer Labeling:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: .. Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. For non-quantitative expression analysis, cDNA was amplified by PCR for 35 cycles at 60°C as annealing temperature and amplified products were visualized after electrophoresis in a 1–2% agarose gel.

    Expressing:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. For non-quantitative expression analysis, cDNA was amplified by PCR for 35 cycles at 60°C as annealing temperature and amplified products were visualized after electrophoresis in a 1–2% agarose gel.

    Article Title: Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells
    Article Snippet: Quantitative real-time PCR (qPCR) cDNA was synthesized using Superscript III Reverse Transcriptase (Sigma) according to the manufacturer’s instructions. qPCR was performed in 96-well plates using iQ™ SYBR Green Supermix (Bio-Rad) and an I-cycler iQ Real-Time PCR instrument (Bio-Rad). .. Measurements were performed in triplicate with a variability < 0.5 Ct. mRNA expression was analysed by normalizing to that of the housekeeping gene GADPH.

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. For mCherry-positive flow cytometer sorted tumour and lung metastatic cells, cDNA was produced directly from lysed cells using a TaqMan Gene Expression Cells-to-Ct Kit (Thermo Fisher Scientific). qPCR was performed on an CFX96 (Bio-Rad) using TaqMan primer/probe sets and all signals were quantified as described above.

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: In brief, HEK293 cells expressing NSUN6-HisPrcFlag, NSUN6-C373A-HisPrcFLAG or the HisPrcFlag tag alone were induced using 1 μg/mL tetracycline for 24 h and UV crosslinking was carried out using a Stratalinker (Stratagene). .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Activated Clotting Time Assay:

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. All signals were quantified using the ACt method and were normalized to the levels of Gapdh.

    Flow Cytometry:

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. For mCherry-positive flow cytometer sorted tumour and lung metastatic cells, cDNA was produced directly from lysed cells using a TaqMan Gene Expression Cells-to-Ct Kit (Thermo Fisher Scientific). qPCR was performed on an CFX96 (Bio-Rad) using TaqMan primer/probe sets and all signals were quantified as described above.

    Sequencing:

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: .. Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer. .. To clone the laccase gene from the Cerrena sp. RSD1, the cDNA of DLac was specifically amplified using the degenerated primer NP2.

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates. ..

    SYBR Green Assay:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. Quantitative PCR was performed with a final 1 in 20 dilution of cDNA, 8 µM primers and Sybr Green MasterMix (Invitrogen).

    Article Title: Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells
    Article Snippet: .. Quantitative real-time PCR (qPCR) cDNA was synthesized using Superscript III Reverse Transcriptase (Sigma) according to the manufacturer’s instructions. qPCR was performed in 96-well plates using iQ™ SYBR Green Supermix (Bio-Rad) and an I-cycler iQ Real-Time PCR instrument (Bio-Rad). .. Measurements were performed in triplicate with a variability < 0.5 Ct. mRNA expression was analysed by normalizing to that of the housekeeping gene GADPH.

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    Cell Culture:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: RNA extraction, reverse transcription, quantitative PCR and PCR Total RNA from cultured cells or tumor tissues was purified using Trizol (Invitrogen) and treated with DNase I (Sigma) according to the manufacturer's instructions. .. Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously .

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: .. Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer. .. To clone the laccase gene from the Cerrena sp. RSD1, the cDNA of DLac was specifically amplified using the degenerated primer NP2.

    Sonication:

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: For chemical crosslinking, cells were incubated in growth media complemented with 5-azacytidine (Sigma), harvested in buffer containing 50 mM Tris/HCl pH 7.6, 150 mM NaCl, 0.1% NP-40, 5 mM β-mercaptoethanol and protease inhibitors, and lysed by sonication. .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Affinity Purification:

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: Protein–RNA complexes were tandem affinity purified using anti-FLAG magnetic beads (Sigma) followed by elution with PreScission Protease. .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Cellular Antioxidant Activity Assay:

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    Nucleic Acid Electrophoresis:

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: The RNA was partially digested (1 min with 1 unit RNace-IT [Agilent]) and radiolabeled using T4 PNK (Thermo Scientific) and [γ-32 P]ATP (PerkinElmer); complexes were purified using Nickel-NTA (Qiagen) under denaturing conditions (6 M guanidium-HCL) and transferred to a nitrocellulose membrane after separation by NuPAGE gel electrophoresis. .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Magnetic Beads:

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: Protein–RNA complexes were tandem affinity purified using anti-FLAG magnetic beads (Sigma) followed by elution with PreScission Protease. .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Isolation:

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: Genomic DNA was extracted using the Wizard® Genomic DNA purification kit (Promega); total RNA was isolated with the TRIzol® Reagent (Ambion®). .. Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer.

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: For whole tissues, RNA was isolated using a TRIzol Plus RNA Purification Kit (Thermo Fisher Scientific). .. RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex.

    Labeling:

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: .. Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer. .. To clone the laccase gene from the Cerrena sp. RSD1, the cDNA of DLac was specifically amplified using the degenerated primer NP2.

    Purification:

    Article Title: A model of breast cancer heterogeneity reveals vascular mimicry as a driver of metastasis
    Article Snippet: Total RNA was purified and DNase treated using the RNeasy Mini Kit (Qiagen). .. Synthesis of cDNA was performed using SuperScript III Reverse Transcriptase (Sigma).

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: RNA extraction, reverse transcription, quantitative PCR and PCR Total RNA from cultured cells or tumor tissues was purified using Trizol (Invitrogen) and treated with DNase I (Sigma) according to the manufacturer's instructions. .. Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously .

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: For whole tissues, RNA was isolated using a TRIzol Plus RNA Purification Kit (Thermo Fisher Scientific). .. RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex.

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: The RNA was partially digested (1 min with 1 unit RNace-IT [Agilent]) and radiolabeled using T4 PNK (Thermo Scientific) and [γ-32 P]ATP (PerkinElmer); complexes were purified using Nickel-NTA (Qiagen) under denaturing conditions (6 M guanidium-HCL) and transferred to a nitrocellulose membrane after separation by NuPAGE gel electrophoresis. .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Polymerase Chain Reaction:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Paragraph title: RNA extraction, reverse transcription, quantitative PCR and PCR ... Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously .

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: TRI reagent (Sigma, St Louis, MO, USA) was immediately added to aliquots of homogenized brain tissues for PCR at 10 times of volume for mRNA extraction. .. Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma).

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer. .. To obtain the signal peptide sequence, 5′‐RACE was performed with SMART II™ A oligonucleotide, the GSP1 primer and 5′ PCR Primer II A.

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates. ..

    cDNA Library Assay:

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates. ..

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    Software:

    Article Title: Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells
    Article Snippet: Quantitative real-time PCR (qPCR) cDNA was synthesized using Superscript III Reverse Transcriptase (Sigma) according to the manufacturer’s instructions. qPCR was performed in 96-well plates using iQ™ SYBR Green Supermix (Bio-Rad) and an I-cycler iQ Real-Time PCR instrument (Bio-Rad). .. Primers were designed with PERL primer software using NCBI EntrezGene reference sequences as templates and synthesized by Sigma.

    Real-time Polymerase Chain Reaction:

    Article Title: A model of breast cancer heterogeneity reveals vascular mimicry as a driver of metastasis
    Article Snippet: Synthesis of cDNA was performed using SuperScript III Reverse Transcriptase (Sigma). .. Quantitative PCR analysis was performed on the Eppendorf Mastercycler ep realplex.

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Paragraph title: RNA extraction, reverse transcription, quantitative PCR and PCR ... Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously .

    Article Title: Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells
    Article Snippet: .. Quantitative real-time PCR (qPCR) cDNA was synthesized using Superscript III Reverse Transcriptase (Sigma) according to the manufacturer’s instructions. qPCR was performed in 96-well plates using iQ™ SYBR Green Supermix (Bio-Rad) and an I-cycler iQ Real-Time PCR instrument (Bio-Rad). .. Measurements were performed in triplicate with a variability < 0.5 Ct. mRNA expression was analysed by normalizing to that of the housekeeping gene GADPH.

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Paragraph title: Real-time PCR ... Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma).

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: .. RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. All signals were quantified using the ACt method and were normalized to the levels of Gapdh.

    RNA Extraction:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Paragraph title: RNA extraction, reverse transcription, quantitative PCR and PCR ... Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously .

    Agarose Gel Electrophoresis:

    Article Title: Emerging Role of Long Non-Coding RNA SOX2OT in SOX2 Regulation in Breast Cancer
    Article Snippet: Oligo-dT or random hexamer were used to reverse transcribe 1 µg of RNA with SuperScript III Reverse Transcriptase (Sigma) according to the manufacturer's instructions. cDNA preparation and qRT-PCR analysis were performed as described previously . .. For non-quantitative expression analysis, cDNA was amplified by PCR for 35 cycles at 60°C as annealing temperature and amplified products were visualized after electrophoresis in a 1–2% agarose gel.

    Incubation:

    Article Title: NSUN6 is a human RNA methyltransferase that catalyzes formation of m5C72 in specific tRNAs
    Article Snippet: For chemical crosslinking, cells were incubated in growth media complemented with 5-azacytidine (Sigma), harvested in buffer containing 50 mM Tris/HCl pH 7.6, 150 mM NaCl, 0.1% NP-40, 5 mM β-mercaptoethanol and protease inhibitors, and lysed by sonication. .. For Illumina deep sequencing, 3′ and 5′ adapters were ligated to the coimmunoprecipitated RNA, the RNA was reverse transcribed using Superscript III Reverse Transcriptase (Sigma) and the cDNA library was amplified using primers that contained randomized sequences of 5 nt to distinguish PCR templates.

    Produced:

    Article Title: Asparagine bioavailability governs metastasis in a model of breast cancer
    Article Snippet: RNA integrity (RNA integrity score > 9) was measured on an Agilent Bioanalyzer (RNA nano kit). cDNA was synthesized using SuperScript III Reverse Transcriptase (Sigma). qPCR was performed on an Eppendorf Mastercycler ep realplex. .. For mCherry-positive flow cytometer sorted tumour and lung metastatic cells, cDNA was produced directly from lysed cells using a TaqMan Gene Expression Cells-to-Ct Kit (Thermo Fisher Scientific). qPCR was performed on an CFX96 (Bio-Rad) using TaqMan primer/probe sets and all signals were quantified as described above.

    Concentration Assay:

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: .. Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

    DNA Purification:

    Article Title: Kinetic analysis and structural studies of a high‐efficiency laccase from Cerrena sp. RSD1
    Article Snippet: Genomic DNA was extracted using the Wizard® Genomic DNA purification kit (Promega); total RNA was isolated with the TRIzol® Reagent (Ambion®). .. Total cDNA was labeled with an anchor sequence by RT‐PCR using SuperScript® III Reverse Transcriptase (Novagen) and the RT‐polyT primer.

    CTG Assay:

    Article Title: Nociceptin/orphanin FQ peptide receptor antagonist JTC-801 reverses pain and anxiety symptoms in a rat model of post-traumatic stress disorder
    Article Snippet: Samples with RNA Quality Index greater than 7 were further examined. cDNA was synthesized to a final concentration of 1 µg µL−1 using SuperScript III Reverse Transcriptase (Sigma). .. Real-time PCR was performed using SYBR Green Master Mix (AnaSpec, Fremont, CA, USA) and 125 nM forward and reverse primers (rat NOP Fwd: 5′-GTT CAA GGA CTG GGT GTT CAG CCA GGT AGT-3′; rat NOP Rev: 5′-TGC TGG CCG TGG TAC TGT CTC AGA ACT CTT-3′; rat GAPDH Fwd: 5′- TGC ACC ACC AAC TGC TTA GC-3′; rat GAPDH Rev: 5′-GGC ATG GAC TGT GGT CAT GAG-3′) and performed in ABI 7000 Sequence Detection System (Applied Biosystems, Foster City, CA, USA).

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  • 93
    Millipore heparitinase
    Enzymatic cleavage of heparan sulfate prevents LTP but has no effect on single pulse-evoked synaptic responses in the area CA1 of hippocampal slices. A , Effect of HFS on the fEPSP slope in rat hippocampal slices (300 μm) preincubated with <t>heparitinase–0.2%</t> BSA (20 U/ml; volume, 500 μl; 3 hr; +24°C) (•) or 0.2% BSA only (○) (average ± SEM; n = 7 on both groups; p
    Heparitinase, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/heparitinase/product/Millipore
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    heparitinase - by Bioz Stars, 2020-04
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    Enzymatic cleavage of heparan sulfate prevents LTP but has no effect on single pulse-evoked synaptic responses in the area CA1 of hippocampal slices. A , Effect of HFS on the fEPSP slope in rat hippocampal slices (300 μm) preincubated with heparitinase–0.2% BSA (20 U/ml; volume, 500 μl; 3 hr; +24°C) (•) or 0.2% BSA only (○) (average ± SEM; n = 7 on both groups; p

    Journal: The Journal of Neuroscience

    Article Title: Reg1ulatory Role and Molecular Interactions of a Cell-Surface Heparan Sulfate Proteoglycan (N-syndecan) in Hippocampal Long-Term Potentiation

    doi: 10.1523/JNEUROSCI.19-04-01226.1999

    Figure Lengend Snippet: Enzymatic cleavage of heparan sulfate prevents LTP but has no effect on single pulse-evoked synaptic responses in the area CA1 of hippocampal slices. A , Effect of HFS on the fEPSP slope in rat hippocampal slices (300 μm) preincubated with heparitinase–0.2% BSA (20 U/ml; volume, 500 μl; 3 hr; +24°C) (•) or 0.2% BSA only (○) (average ± SEM; n = 7 on both groups; p

    Article Snippet: Aliquots of the fractions were digested with nitrous acid or heparitinase (10 U/ml, 15 hr) to remove carbohydrate side chains or incubated with 50 μ m herbimycin A (Calbiochem, La Jolla, CA).

    Techniques: