real time quantitative reverse transcription pcr rt qpcr reactions  (Roche)


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    Roche real time quantitative reverse transcription pcr rt qpcr reactions
    Quantitative <t>RT-PCR</t> analysis of candidate genes. Mock infected plants were used to normalize fold change of early and late samples in leaf and tuber tissues as indicated. White bars represent the <t>RT-qPCR</t> analysis; grey bars show the microarray data. Error bars represent standard error (SE). L23 and PP2A genes were used as internal normalization controls. Asterisk indicates that the means of fold change of the mock and infected samples are significantly different (Student’s t-test).
    Real Time Quantitative Reverse Transcription Pcr Rt Qpcr Reactions, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time quantitative reverse transcription pcr rt qpcr reactions/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    real time quantitative reverse transcription pcr rt qpcr reactions - by Bioz Stars, 2021-07
    86/100 stars

    Images

    1) Product Images from "Insight on Genes Affecting Tuber Development in Potato upon Potato spindle tuber viroid (PSTVd) Infection"

    Article Title: Insight on Genes Affecting Tuber Development in Potato upon Potato spindle tuber viroid (PSTVd) Infection

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0150711

    Quantitative RT-PCR analysis of candidate genes. Mock infected plants were used to normalize fold change of early and late samples in leaf and tuber tissues as indicated. White bars represent the RT-qPCR analysis; grey bars show the microarray data. Error bars represent standard error (SE). L23 and PP2A genes were used as internal normalization controls. Asterisk indicates that the means of fold change of the mock and infected samples are significantly different (Student’s t-test).
    Figure Legend Snippet: Quantitative RT-PCR analysis of candidate genes. Mock infected plants were used to normalize fold change of early and late samples in leaf and tuber tissues as indicated. White bars represent the RT-qPCR analysis; grey bars show the microarray data. Error bars represent standard error (SE). L23 and PP2A genes were used as internal normalization controls. Asterisk indicates that the means of fold change of the mock and infected samples are significantly different (Student’s t-test).

    Techniques Used: Quantitative RT-PCR, Infection, Microarray

    Related Articles

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Piperlongumine inhibits proliferation and survival of Burkitt lymphoma in vitro
    Article Snippet: Reverse transcription (RT) PCR and quantitative PCR (qPCR) relied on: total RNA extracted with the help of the TRIzol (Sigma-Aldrich, St. Louis, MO) reagent; cDNA synthesis using 1 μg RNA and the AMV reverse transcriptase kit from Roche (Indianapolis, IN); and the gene-specific PCR primers listed in . .. RT-PCR amplification of LMP-1 (human herpsesvirus 4), Myc (c-myc) and Actb (b-actin) message used the PCR master mix from Roche and 20–40 cycles of DNA amplification under the following cycling conditions: template denaturation at 95 °C for 5 minutes, primer annealing at 57 °C for 1 minute, and primer extension at 72 °C for 1 minute. qPCR relied on the TaqMan universal PCR master mix from Applied Biosystems (Carlsbad, CA) and – after 1 cycle of 50 C for 2 min and 1 cycle of 95 C for 10 min – 40 cycles of DNA amplification at 66 °C (1 min) and 95 °C (15 sec). .. Internal probes were labeled with the fluorescent reporter dye, 6-carboxyfluorescein (6- FAM), on the 5′ end and the quencher dye, Black Hole (BHQ), on the 3′ end.

    Amplification:

    Article Title: Piperlongumine inhibits proliferation and survival of Burkitt lymphoma in vitro
    Article Snippet: Reverse transcription (RT) PCR and quantitative PCR (qPCR) relied on: total RNA extracted with the help of the TRIzol (Sigma-Aldrich, St. Louis, MO) reagent; cDNA synthesis using 1 μg RNA and the AMV reverse transcriptase kit from Roche (Indianapolis, IN); and the gene-specific PCR primers listed in . .. RT-PCR amplification of LMP-1 (human herpsesvirus 4), Myc (c-myc) and Actb (b-actin) message used the PCR master mix from Roche and 20–40 cycles of DNA amplification under the following cycling conditions: template denaturation at 95 °C for 5 minutes, primer annealing at 57 °C for 1 minute, and primer extension at 72 °C for 1 minute. qPCR relied on the TaqMan universal PCR master mix from Applied Biosystems (Carlsbad, CA) and – after 1 cycle of 50 C for 2 min and 1 cycle of 95 C for 10 min – 40 cycles of DNA amplification at 66 °C (1 min) and 95 °C (15 sec). .. Internal probes were labeled with the fluorescent reporter dye, 6-carboxyfluorescein (6- FAM), on the 5′ end and the quencher dye, Black Hole (BHQ), on the 3′ end.

    Polymerase Chain Reaction:

    Article Title: Piperlongumine inhibits proliferation and survival of Burkitt lymphoma in vitro
    Article Snippet: Reverse transcription (RT) PCR and quantitative PCR (qPCR) relied on: total RNA extracted with the help of the TRIzol (Sigma-Aldrich, St. Louis, MO) reagent; cDNA synthesis using 1 μg RNA and the AMV reverse transcriptase kit from Roche (Indianapolis, IN); and the gene-specific PCR primers listed in . .. RT-PCR amplification of LMP-1 (human herpsesvirus 4), Myc (c-myc) and Actb (b-actin) message used the PCR master mix from Roche and 20–40 cycles of DNA amplification under the following cycling conditions: template denaturation at 95 °C for 5 minutes, primer annealing at 57 °C for 1 minute, and primer extension at 72 °C for 1 minute. qPCR relied on the TaqMan universal PCR master mix from Applied Biosystems (Carlsbad, CA) and – after 1 cycle of 50 C for 2 min and 1 cycle of 95 C for 10 min – 40 cycles of DNA amplification at 66 °C (1 min) and 95 °C (15 sec). .. Internal probes were labeled with the fluorescent reporter dye, 6-carboxyfluorescein (6- FAM), on the 5′ end and the quencher dye, Black Hole (BHQ), on the 3′ end.

    Article Title: In Vivo Excision of HIV-1 Provirus by saCas9 and Multiplex Single-Guide RNAs in Animal Models
    Article Snippet: The potentially residual genomic DNA was removed through on-column DNase digestion with an RNase-Free DNase Set (QIAGEN). .. One μg of RNA for each sample was reverse transcribed into cDNAs using random hexanucleotide primers with a High-Capacity cDNA Reverse Transcription Kit (Invitrogen). qPCR analyses of cDNA or HIV genomic DNA were carried out in a LightCycler480 (Roche) using a SYBR Green PCR Master Mix Kit (Applied Biosystems). ..

    Article Title: Activated leukocyte cell adhesion molecule regulates the interaction between pancreatic cancer cells and stellate cells
    Article Snippet: PSCs between passages 3 and 6 were cultured in a 1:1 (vol/vol) mixture of low-glucose (1,000 mg/l) Dulbecco's modified Eagle's medium with Ham's F12 medium (Invitrogen; Thermo Fisher Scientific, Inc.) supplemented with 10% FCS, L-glutamine (2 mmol/l), penicillin/streptomycin and amphotericin, as previously described ( – ). .. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) All reagents and equipment for mRNA and cDNA preparation were purchased from Roche (Mannheim, Germany). .. The forward and reverse primer sequences were as follows: ALCAM sense, 5′-TAG CAG GAA TGC AAC TGT GG-3′; ALCAM anti-sense, 5′-CGC AGA CAT AGT TTC CAG-3′. mRNA was prepared by automated isolation using the MagNA Pure LC instrument and isolation kit I (for cells) and kit II (for tissues).

    Article Title: Genome-wide mapping of histone H3K9me2 in acute myeloid leukemia reveals large chromosomal domains associated with massive gene silencing and sites of genome instability
    Article Snippet: First strand cDNA was produced from 1.0 μg of total RNA using the standard protocol from the High Capacity cDNA Reverse Transcriptase kit (Life Technologies, Grand Island, NY). cDNA concentrations were quantified by absorbance using a NanoDrop ND-1000 Spectrophotometer (Fisher Scientific, Pittsburgh, PA). cDNA expression rates in AML were assayed using Illumina HumanHT-12 array containing hybridization probes with 33,732 distinct chromosomal positions and using normal bone marrow CD34+ cells as controls (see Table D in ). .. Genomic qPCR, RT-qPCR and PCR primers All quantitative real-time PCR (qPCR) was performed using FastStart Universal SYBR Green Master (Roche) on a Roche LightCycler 480 Instrument following the manufacturer’s instructions. .. For ChIP-qPCR assays, reactions were performed in triplicate, with two technical replicates for each sample.

    Real-time Polymerase Chain Reaction:

    Article Title: Piperlongumine inhibits proliferation and survival of Burkitt lymphoma in vitro
    Article Snippet: Reverse transcription (RT) PCR and quantitative PCR (qPCR) relied on: total RNA extracted with the help of the TRIzol (Sigma-Aldrich, St. Louis, MO) reagent; cDNA synthesis using 1 μg RNA and the AMV reverse transcriptase kit from Roche (Indianapolis, IN); and the gene-specific PCR primers listed in . .. RT-PCR amplification of LMP-1 (human herpsesvirus 4), Myc (c-myc) and Actb (b-actin) message used the PCR master mix from Roche and 20–40 cycles of DNA amplification under the following cycling conditions: template denaturation at 95 °C for 5 minutes, primer annealing at 57 °C for 1 minute, and primer extension at 72 °C for 1 minute. qPCR relied on the TaqMan universal PCR master mix from Applied Biosystems (Carlsbad, CA) and – after 1 cycle of 50 C for 2 min and 1 cycle of 95 C for 10 min – 40 cycles of DNA amplification at 66 °C (1 min) and 95 °C (15 sec). .. Internal probes were labeled with the fluorescent reporter dye, 6-carboxyfluorescein (6- FAM), on the 5′ end and the quencher dye, Black Hole (BHQ), on the 3′ end.

    Article Title: Mesenchymal Stem Cells Ameliorated Glucolipotoxicity in HUVECs through TSG-6
    Article Snippet: Cell apoptosis was also analyzed via flow cytometry by Annexin-V-FLUOS Staining Kit (Roche) following the manufacture’s protocol. .. Total RNA Extraction and Real-Time qPCR MSCs, HSFs and HUVECs were stimulated with 30 mM glucose and 100 μM palmitic acid for 24 h. Then cell (approximately 3 × 105 cells per sample) RNA was extracted via Tripure Isolation Reagent (Roche). .. The cDNA was synthesized by reverse transcription using Transcriptor First Strand cDNA Synthesis Kit (Roche).

    Article Title: The histone deacetylase inhibitor PCI-24781 impairs calcium influx and inhibits proliferation and metastasis in breast cancer
    Article Snippet: All RNA-seq data are available on https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE150038. .. Real-time quantitative PCR analysis For quantification of mRNA of target genes by RT-qPCR, total RNA was extracted from MDA-MB-231 cells treated with PCI-24781 using a TRIzol reagent (Roche, Basel, Switzerland). .. Reverse transcription was performed using the RevertAid First Strand cDNA Synthesis Kit (Roche, Basel, Switzerland) according to the manufacturer's instructions.

    Article Title: In Vivo Excision of HIV-1 Provirus by saCas9 and Multiplex Single-Guide RNAs in Animal Models
    Article Snippet: The potentially residual genomic DNA was removed through on-column DNase digestion with an RNase-Free DNase Set (QIAGEN). .. One μg of RNA for each sample was reverse transcribed into cDNAs using random hexanucleotide primers with a High-Capacity cDNA Reverse Transcription Kit (Invitrogen). qPCR analyses of cDNA or HIV genomic DNA were carried out in a LightCycler480 (Roche) using a SYBR Green PCR Master Mix Kit (Applied Biosystems). ..

    Article Title: Genome-wide mapping of histone H3K9me2 in acute myeloid leukemia reveals large chromosomal domains associated with massive gene silencing and sites of genome instability
    Article Snippet: First strand cDNA was produced from 1.0 μg of total RNA using the standard protocol from the High Capacity cDNA Reverse Transcriptase kit (Life Technologies, Grand Island, NY). cDNA concentrations were quantified by absorbance using a NanoDrop ND-1000 Spectrophotometer (Fisher Scientific, Pittsburgh, PA). cDNA expression rates in AML were assayed using Illumina HumanHT-12 array containing hybridization probes with 33,732 distinct chromosomal positions and using normal bone marrow CD34+ cells as controls (see Table D in ). .. Genomic qPCR, RT-qPCR and PCR primers All quantitative real-time PCR (qPCR) was performed using FastStart Universal SYBR Green Master (Roche) on a Roche LightCycler 480 Instrument following the manufacturer’s instructions. .. For ChIP-qPCR assays, reactions were performed in triplicate, with two technical replicates for each sample.

    Article Title: Rheumatoid Arthritis-Associated MicroRNA-155 Targets SOCS1 and Upregulates TNF-? and IL-1? in PBMCs
    Article Snippet: RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) Total RNA was extracted from tissue samples by using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). .. Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) analysis of the TNF-α, IL-1β and SOCS1 mRNA in PBMCs was performed using SYBR Green with the LightCycle 2.0 (Roche, Mannheim, Germany). .. All RNA expression levels were normalized to GAPDH (glyceraldehyde-3-phosphate dehydrogenase). miRNA extraction from PBMCs was performed by using the mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA).

    Article Title: MiR-519d represses ovarian cancer cell proliferation and enhances cisplatin-mediated cytotoxicity in vitro by targeting XIAP
    Article Snippet: RNA extraction and quantitative real-time PCR analyses of mRNA Total RNAs were isolated using Trizol (Invitrogen Life Technologies), according to the manufacturer’s instructions. .. One microgram total RNA was used for reverse transcription, using a ReverTra Ace qPCR-RT Kit (Toyobo, Osaka, Japan) in a reaction volume of 10 μL. qRT-PCR was performed to detect the relative level of XIAP mRNA, using the Light-Cycler rapid thermal cycler system 2.0 (Roche Diagnostics Ltd, Burgess Hill, United Kingdom) with SYBR Green Master Mix (Toyobo). .. Primers used for real-time PCR are as follows: XIAP (forward) 5′-CCGTGCGGTGCTTTAGTTGT-3′ (reverse) 5′-TTCCTCGGGTATATGGTGTCTGAT-3′; GAPDH (forward) 5′-TGGTCACCAGGGCTGCTT-3′ (reverse) 5′-AGCTTCCCGTTCTCAGCCTT-3′.

    Size-exclusion Chromatography:

    Article Title: Piperlongumine inhibits proliferation and survival of Burkitt lymphoma in vitro
    Article Snippet: Reverse transcription (RT) PCR and quantitative PCR (qPCR) relied on: total RNA extracted with the help of the TRIzol (Sigma-Aldrich, St. Louis, MO) reagent; cDNA synthesis using 1 μg RNA and the AMV reverse transcriptase kit from Roche (Indianapolis, IN); and the gene-specific PCR primers listed in . .. RT-PCR amplification of LMP-1 (human herpsesvirus 4), Myc (c-myc) and Actb (b-actin) message used the PCR master mix from Roche and 20–40 cycles of DNA amplification under the following cycling conditions: template denaturation at 95 °C for 5 minutes, primer annealing at 57 °C for 1 minute, and primer extension at 72 °C for 1 minute. qPCR relied on the TaqMan universal PCR master mix from Applied Biosystems (Carlsbad, CA) and – after 1 cycle of 50 C for 2 min and 1 cycle of 95 C for 10 min – 40 cycles of DNA amplification at 66 °C (1 min) and 95 °C (15 sec). .. Internal probes were labeled with the fluorescent reporter dye, 6-carboxyfluorescein (6- FAM), on the 5′ end and the quencher dye, Black Hole (BHQ), on the 3′ end.

    RNA Extraction:

    Article Title: Mesenchymal Stem Cells Ameliorated Glucolipotoxicity in HUVECs through TSG-6
    Article Snippet: Cell apoptosis was also analyzed via flow cytometry by Annexin-V-FLUOS Staining Kit (Roche) following the manufacture’s protocol. .. Total RNA Extraction and Real-Time qPCR MSCs, HSFs and HUVECs were stimulated with 30 mM glucose and 100 μM palmitic acid for 24 h. Then cell (approximately 3 × 105 cells per sample) RNA was extracted via Tripure Isolation Reagent (Roche). .. The cDNA was synthesized by reverse transcription using Transcriptor First Strand cDNA Synthesis Kit (Roche).

    Isolation:

    Article Title: Mesenchymal Stem Cells Ameliorated Glucolipotoxicity in HUVECs through TSG-6
    Article Snippet: Cell apoptosis was also analyzed via flow cytometry by Annexin-V-FLUOS Staining Kit (Roche) following the manufacture’s protocol. .. Total RNA Extraction and Real-Time qPCR MSCs, HSFs and HUVECs were stimulated with 30 mM glucose and 100 μM palmitic acid for 24 h. Then cell (approximately 3 × 105 cells per sample) RNA was extracted via Tripure Isolation Reagent (Roche). .. The cDNA was synthesized by reverse transcription using Transcriptor First Strand cDNA Synthesis Kit (Roche).

    Quantitative RT-PCR:

    Article Title: The histone deacetylase inhibitor PCI-24781 impairs calcium influx and inhibits proliferation and metastasis in breast cancer
    Article Snippet: All RNA-seq data are available on https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE150038. .. Real-time quantitative PCR analysis For quantification of mRNA of target genes by RT-qPCR, total RNA was extracted from MDA-MB-231 cells treated with PCI-24781 using a TRIzol reagent (Roche, Basel, Switzerland). .. Reverse transcription was performed using the RevertAid First Strand cDNA Synthesis Kit (Roche, Basel, Switzerland) according to the manufacturer's instructions.

    Article Title: Activated leukocyte cell adhesion molecule regulates the interaction between pancreatic cancer cells and stellate cells
    Article Snippet: PSCs between passages 3 and 6 were cultured in a 1:1 (vol/vol) mixture of low-glucose (1,000 mg/l) Dulbecco's modified Eagle's medium with Ham's F12 medium (Invitrogen; Thermo Fisher Scientific, Inc.) supplemented with 10% FCS, L-glutamine (2 mmol/l), penicillin/streptomycin and amphotericin, as previously described ( – ). .. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) All reagents and equipment for mRNA and cDNA preparation were purchased from Roche (Mannheim, Germany). .. The forward and reverse primer sequences were as follows: ALCAM sense, 5′-TAG CAG GAA TGC AAC TGT GG-3′; ALCAM anti-sense, 5′-CGC AGA CAT AGT TTC CAG-3′. mRNA was prepared by automated isolation using the MagNA Pure LC instrument and isolation kit I (for cells) and kit II (for tissues).

    Article Title: Genome-wide mapping of histone H3K9me2 in acute myeloid leukemia reveals large chromosomal domains associated with massive gene silencing and sites of genome instability
    Article Snippet: First strand cDNA was produced from 1.0 μg of total RNA using the standard protocol from the High Capacity cDNA Reverse Transcriptase kit (Life Technologies, Grand Island, NY). cDNA concentrations were quantified by absorbance using a NanoDrop ND-1000 Spectrophotometer (Fisher Scientific, Pittsburgh, PA). cDNA expression rates in AML were assayed using Illumina HumanHT-12 array containing hybridization probes with 33,732 distinct chromosomal positions and using normal bone marrow CD34+ cells as controls (see Table D in ). .. Genomic qPCR, RT-qPCR and PCR primers All quantitative real-time PCR (qPCR) was performed using FastStart Universal SYBR Green Master (Roche) on a Roche LightCycler 480 Instrument following the manufacturer’s instructions. .. For ChIP-qPCR assays, reactions were performed in triplicate, with two technical replicates for each sample.

    Article Title: Rheumatoid Arthritis-Associated MicroRNA-155 Targets SOCS1 and Upregulates TNF-? and IL-1? in PBMCs
    Article Snippet: RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) Total RNA was extracted from tissue samples by using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). .. Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) analysis of the TNF-α, IL-1β and SOCS1 mRNA in PBMCs was performed using SYBR Green with the LightCycle 2.0 (Roche, Mannheim, Germany). .. All RNA expression levels were normalized to GAPDH (glyceraldehyde-3-phosphate dehydrogenase). miRNA extraction from PBMCs was performed by using the mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA).

    Article Title: MiR-519d represses ovarian cancer cell proliferation and enhances cisplatin-mediated cytotoxicity in vitro by targeting XIAP
    Article Snippet: RNA extraction and quantitative real-time PCR analyses of mRNA Total RNAs were isolated using Trizol (Invitrogen Life Technologies), according to the manufacturer’s instructions. .. One microgram total RNA was used for reverse transcription, using a ReverTra Ace qPCR-RT Kit (Toyobo, Osaka, Japan) in a reaction volume of 10 μL. qRT-PCR was performed to detect the relative level of XIAP mRNA, using the Light-Cycler rapid thermal cycler system 2.0 (Roche Diagnostics Ltd, Burgess Hill, United Kingdom) with SYBR Green Master Mix (Toyobo). .. Primers used for real-time PCR are as follows: XIAP (forward) 5′-CCGTGCGGTGCTTTAGTTGT-3′ (reverse) 5′-TTCCTCGGGTATATGGTGTCTGAT-3′; GAPDH (forward) 5′-TGGTCACCAGGGCTGCTT-3′ (reverse) 5′-AGCTTCCCGTTCTCAGCCTT-3′.

    Multiple Displacement Amplification:

    Article Title: The histone deacetylase inhibitor PCI-24781 impairs calcium influx and inhibits proliferation and metastasis in breast cancer
    Article Snippet: All RNA-seq data are available on https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE150038. .. Real-time quantitative PCR analysis For quantification of mRNA of target genes by RT-qPCR, total RNA was extracted from MDA-MB-231 cells treated with PCI-24781 using a TRIzol reagent (Roche, Basel, Switzerland). .. Reverse transcription was performed using the RevertAid First Strand cDNA Synthesis Kit (Roche, Basel, Switzerland) according to the manufacturer's instructions.

    SYBR Green Assay:

    Article Title: In Vivo Excision of HIV-1 Provirus by saCas9 and Multiplex Single-Guide RNAs in Animal Models
    Article Snippet: The potentially residual genomic DNA was removed through on-column DNase digestion with an RNase-Free DNase Set (QIAGEN). .. One μg of RNA for each sample was reverse transcribed into cDNAs using random hexanucleotide primers with a High-Capacity cDNA Reverse Transcription Kit (Invitrogen). qPCR analyses of cDNA or HIV genomic DNA were carried out in a LightCycler480 (Roche) using a SYBR Green PCR Master Mix Kit (Applied Biosystems). ..

    Article Title: Genome-wide mapping of histone H3K9me2 in acute myeloid leukemia reveals large chromosomal domains associated with massive gene silencing and sites of genome instability
    Article Snippet: First strand cDNA was produced from 1.0 μg of total RNA using the standard protocol from the High Capacity cDNA Reverse Transcriptase kit (Life Technologies, Grand Island, NY). cDNA concentrations were quantified by absorbance using a NanoDrop ND-1000 Spectrophotometer (Fisher Scientific, Pittsburgh, PA). cDNA expression rates in AML were assayed using Illumina HumanHT-12 array containing hybridization probes with 33,732 distinct chromosomal positions and using normal bone marrow CD34+ cells as controls (see Table D in ). .. Genomic qPCR, RT-qPCR and PCR primers All quantitative real-time PCR (qPCR) was performed using FastStart Universal SYBR Green Master (Roche) on a Roche LightCycler 480 Instrument following the manufacturer’s instructions. .. For ChIP-qPCR assays, reactions were performed in triplicate, with two technical replicates for each sample.

    Article Title: Rheumatoid Arthritis-Associated MicroRNA-155 Targets SOCS1 and Upregulates TNF-? and IL-1? in PBMCs
    Article Snippet: RNA Extraction and Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) Total RNA was extracted from tissue samples by using the RNeasy Mini Kit (Qiagen, Valencia, CA, USA). .. Reverse-transcription quantitative polymerase chain reaction (RT-qPCR) analysis of the TNF-α, IL-1β and SOCS1 mRNA in PBMCs was performed using SYBR Green with the LightCycle 2.0 (Roche, Mannheim, Germany). .. All RNA expression levels were normalized to GAPDH (glyceraldehyde-3-phosphate dehydrogenase). miRNA extraction from PBMCs was performed by using the mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA).

    Article Title: MiR-519d represses ovarian cancer cell proliferation and enhances cisplatin-mediated cytotoxicity in vitro by targeting XIAP
    Article Snippet: RNA extraction and quantitative real-time PCR analyses of mRNA Total RNAs were isolated using Trizol (Invitrogen Life Technologies), according to the manufacturer’s instructions. .. One microgram total RNA was used for reverse transcription, using a ReverTra Ace qPCR-RT Kit (Toyobo, Osaka, Japan) in a reaction volume of 10 μL. qRT-PCR was performed to detect the relative level of XIAP mRNA, using the Light-Cycler rapid thermal cycler system 2.0 (Roche Diagnostics Ltd, Burgess Hill, United Kingdom) with SYBR Green Master Mix (Toyobo). .. Primers used for real-time PCR are as follows: XIAP (forward) 5′-CCGTGCGGTGCTTTAGTTGT-3′ (reverse) 5′-TTCCTCGGGTATATGGTGTCTGAT-3′; GAPDH (forward) 5′-TGGTCACCAGGGCTGCTT-3′ (reverse) 5′-AGCTTCCCGTTCTCAGCCTT-3′.

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    Roche reverse transcription quantitative polymerase chain reaction rt qpcr
    Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM <t>mRNA</t> levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l cDNA. Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.
    Reverse Transcription Quantitative Polymerase Chain Reaction Rt Qpcr, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/reverse transcription quantitative polymerase chain reaction rt qpcr/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    reverse transcription quantitative polymerase chain reaction rt qpcr - by Bioz Stars, 2021-07
    86/100 stars
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    86
    Roche real time pcr qpcr
    Inhibition of G9a/GLP in K562 decreases H3K9me2 levels and activates genes within AML-enriched transient H3K9me2 blocks. A: Western blotting probed with antibodies against H3K9me2 and unmodified histone H3 show decreasing levels of H3K9me2 in K562 cells treated with G9a/GLP inhibitor UNC0638. B: Graphs showing correlations between H3K9me2 domains in control and 1 μM UNC0638-treated K562 cells vs. selected biodata with top discriminatory power (insulator, repressed chromatin, H3K27me3, H3K9me3, and heterochromatin). C: Ingenuity pathway analysis showing upstream regulators associated with the dLOCK UNC0638 > K562control and dLOCK K562control > UNC0638 . D: H3K9me2 levels determined by <t>ChIP-qPCR</t> in control and UNC0638-treated K562 cells for selected gene probes representing dLOCK CD34+ > AML ( ZNF274 , ZNF544 ), constitutively high H3K9me2 ( NLRP11 ), constitutively low H3K9me2 ( TRIM28 ), dLOCK AML > CD34+ ( MECOM , ETS1 , ERG ) and dLOCK AML > Gran ( CDH1 ). E: <t>RT-PCR</t> analysis of gene expression level in control and UNC0638-treated K562 cells for selected gene probes representing dLOCK CD34+ > AML ( ZNF274 , ZNF544 ), constitutively low H3K9me2 ( TRIM28 ), dLOCK AML > CD34+ ( MECOM , ETS1 , ERG ) and dLOCK AML > Gran ( CDH1 ). p– values represent Student’s t-test for 2 tailed, unpaired equal variance.
    Real Time Pcr Qpcr, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr qpcr/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    real time pcr qpcr - by Bioz Stars, 2021-07
    86/100 stars
      Buy from Supplier

    86
    Roche quantitative real time rt pcr analysis quantitative real time rt pcr qpcr
    <t>qPCR</t> analysis of differentially expressed genes in Poncirus trifoliata under low temperature. ( a , b ) Transcript levels of 17 randomly selected DEGs, including 12 up-regulated ( a ) and 5 down-regulated ( b ). The Y-axis on the left shows the relative gene expression levels analyzed by qPCR (red lines), while Y-axis on the right shows corresponding expression data of RNA-seq (gray histogram). The X-axis represents the time (hours) of 4 °C treatment. The bars represent SE (n = 3). ( c ) Comparison between the log 2 of gene expression ratios obtained from RNA-seq data and <t>qRT-PCR</t>
    Quantitative Real Time Rt Pcr Analysis Quantitative Real Time Rt Pcr Qpcr, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time rt pcr analysis quantitative real time rt pcr qpcr/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    quantitative real time rt pcr analysis quantitative real time rt pcr qpcr - by Bioz Stars, 2021-07
    86/100 stars
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    86
    Roche quantitative real time pcr qpcr
    AUF-1 facilitates ELAVL4-mediated APP695-specific AS. Biotinylated RNA probes transcribed from human ( A ) and mouse ( B ) APP minigenes were tested for AUF-1 binding. Note, that AUF-1 isoforms p42 and/or p45 bind to all transcripts shown to interact with nELAVLs. ( C ) SK-N-SH cells were transfected with the DNA3.1 expression vector bearing either no insert (empty) or p42 AUF-1, or with the pENTR/U6 vector carrying a shRNA specific for LacZ or all AUF - 1 mRNAs. Inclusion of APP exons 7 and 8 was assayed by <t>RT-qPCR</t> with primers specific for APP770, APP695 and APP751 . Total APP cDNA was used for normalization. Bars in graphs correspond to mean ± standard deviation of three independent experiments. ( D , E ) SK-N-SH cells were co-transfected with the human ( D ) or the mouse ( E ) APPE78 minigene along with two expression vectors, one bearing ELAVL4 and the other one carrying no insert (empty) or p42 AUF-1, in ratios depicted. Splicing pathways were determined by <t>RT-PCR,</t> as described in Fig. 5 . Quantification of the results was performed by scanning densitometry. Note, that AUF-1 enhances ELAVL4-mediated exclusion of APP exons 7 and 8. ( F ) Neuro2a cells were transfected with the pSilencer vector bearing either no insert (empty) or a shRNA targeting all Auf - 1 transcripts. Inclusion of APP exons 7 and 8 was assayed by RT-qPCR, as described above (**P
    Quantitative Real Time Pcr Qpcr, supplied by Roche, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr qpcr/product/Roche
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr qpcr - by Bioz Stars, 2021-07
    86/100 stars
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    Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM mRNA levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l cDNA. Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.

    Journal: Molecular Medicine Reports

    Article Title: Activated leukocyte cell adhesion molecule regulates the interaction between pancreatic cancer cells and stellate cells

    doi: 10.3892/mmr.2016.5681

    Figure Lengend Snippet: Differential expression of ALCAM in pancreatic cancer cells and PSCs. (A) Reverse transcription-quantitative polymerase chain reaction analysis of ALCAM mRNA levels in the pancreatic cancer cell lines Panc-1 and T3M4, as well as in PSCs. RNA input was normalized against the average expression of hypoxanthine-guanine phosphoribosyltransferase and cylophilin B, and presented as the copy number/ µ l cDNA. Immunoblotting analysis was performed to detect ALCAM protein expression in Panc-1 and T3M4 cells, and also in PSCs. (B) Equal loading of the protein samples was confirmed using an ERK-2 antibody. ALCAM, activated leukocyte cell adhesion molecule; PSCs, pancreatic stellate cells; ERK-2, extracellular signal-regulated kinase-2.

    Article Snippet: Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) All reagents and equipment for mRNA and cDNA preparation were purchased from Roche (Mannheim, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction

    Inhibition of G9a/GLP in K562 decreases H3K9me2 levels and activates genes within AML-enriched transient H3K9me2 blocks. A: Western blotting probed with antibodies against H3K9me2 and unmodified histone H3 show decreasing levels of H3K9me2 in K562 cells treated with G9a/GLP inhibitor UNC0638. B: Graphs showing correlations between H3K9me2 domains in control and 1 μM UNC0638-treated K562 cells vs. selected biodata with top discriminatory power (insulator, repressed chromatin, H3K27me3, H3K9me3, and heterochromatin). C: Ingenuity pathway analysis showing upstream regulators associated with the dLOCK UNC0638 > K562control and dLOCK K562control > UNC0638 . D: H3K9me2 levels determined by ChIP-qPCR in control and UNC0638-treated K562 cells for selected gene probes representing dLOCK CD34+ > AML ( ZNF274 , ZNF544 ), constitutively high H3K9me2 ( NLRP11 ), constitutively low H3K9me2 ( TRIM28 ), dLOCK AML > CD34+ ( MECOM , ETS1 , ERG ) and dLOCK AML > Gran ( CDH1 ). E: RT-PCR analysis of gene expression level in control and UNC0638-treated K562 cells for selected gene probes representing dLOCK CD34+ > AML ( ZNF274 , ZNF544 ), constitutively low H3K9me2 ( TRIM28 ), dLOCK AML > CD34+ ( MECOM , ETS1 , ERG ) and dLOCK AML > Gran ( CDH1 ). p– values represent Student’s t-test for 2 tailed, unpaired equal variance.

    Journal: PLoS ONE

    Article Title: Genome-wide mapping of histone H3K9me2 in acute myeloid leukemia reveals large chromosomal domains associated with massive gene silencing and sites of genome instability

    doi: 10.1371/journal.pone.0173723

    Figure Lengend Snippet: Inhibition of G9a/GLP in K562 decreases H3K9me2 levels and activates genes within AML-enriched transient H3K9me2 blocks. A: Western blotting probed with antibodies against H3K9me2 and unmodified histone H3 show decreasing levels of H3K9me2 in K562 cells treated with G9a/GLP inhibitor UNC0638. B: Graphs showing correlations between H3K9me2 domains in control and 1 μM UNC0638-treated K562 cells vs. selected biodata with top discriminatory power (insulator, repressed chromatin, H3K27me3, H3K9me3, and heterochromatin). C: Ingenuity pathway analysis showing upstream regulators associated with the dLOCK UNC0638 > K562control and dLOCK K562control > UNC0638 . D: H3K9me2 levels determined by ChIP-qPCR in control and UNC0638-treated K562 cells for selected gene probes representing dLOCK CD34+ > AML ( ZNF274 , ZNF544 ), constitutively high H3K9me2 ( NLRP11 ), constitutively low H3K9me2 ( TRIM28 ), dLOCK AML > CD34+ ( MECOM , ETS1 , ERG ) and dLOCK AML > Gran ( CDH1 ). E: RT-PCR analysis of gene expression level in control and UNC0638-treated K562 cells for selected gene probes representing dLOCK CD34+ > AML ( ZNF274 , ZNF544 ), constitutively low H3K9me2 ( TRIM28 ), dLOCK AML > CD34+ ( MECOM , ETS1 , ERG ) and dLOCK AML > Gran ( CDH1 ). p– values represent Student’s t-test for 2 tailed, unpaired equal variance.

    Article Snippet: Genomic qPCR, RT-qPCR and PCR primers All quantitative real-time PCR (qPCR) was performed using FastStart Universal SYBR Green Master (Roche) on a Roche LightCycler 480 Instrument following the manufacturer’s instructions.

    Techniques: Inhibition, Western Blot, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Reverse Transcription Polymerase Chain Reaction, Expressing

    qPCR analysis of differentially expressed genes in Poncirus trifoliata under low temperature. ( a , b ) Transcript levels of 17 randomly selected DEGs, including 12 up-regulated ( a ) and 5 down-regulated ( b ). The Y-axis on the left shows the relative gene expression levels analyzed by qPCR (red lines), while Y-axis on the right shows corresponding expression data of RNA-seq (gray histogram). The X-axis represents the time (hours) of 4 °C treatment. The bars represent SE (n = 3). ( c ) Comparison between the log 2 of gene expression ratios obtained from RNA-seq data and qRT-PCR

    Journal: BMC Genomics

    Article Title: Deep sequencing-based characterization of transcriptome of trifoliate orange (Poncirus trifoliata (L.) Raf.) in response to cold stress

    doi: 10.1186/s12864-015-1629-7

    Figure Lengend Snippet: qPCR analysis of differentially expressed genes in Poncirus trifoliata under low temperature. ( a , b ) Transcript levels of 17 randomly selected DEGs, including 12 up-regulated ( a ) and 5 down-regulated ( b ). The Y-axis on the left shows the relative gene expression levels analyzed by qPCR (red lines), while Y-axis on the right shows corresponding expression data of RNA-seq (gray histogram). The X-axis represents the time (hours) of 4 °C treatment. The bars represent SE (n = 3). ( c ) Comparison between the log 2 of gene expression ratios obtained from RNA-seq data and qRT-PCR

    Article Snippet: Quantitative real-time RT-PCR analysis Quantitative real-time RT-PCR (qPCR) was performed with a Roche LightCycler 480 Real-Time System (Roche, Switzerland) to examine expression patterns of the selected unigenes.

    Techniques: Real-time Polymerase Chain Reaction, Expressing, RNA Sequencing Assay, Quantitative RT-PCR

    AUF-1 facilitates ELAVL4-mediated APP695-specific AS. Biotinylated RNA probes transcribed from human ( A ) and mouse ( B ) APP minigenes were tested for AUF-1 binding. Note, that AUF-1 isoforms p42 and/or p45 bind to all transcripts shown to interact with nELAVLs. ( C ) SK-N-SH cells were transfected with the DNA3.1 expression vector bearing either no insert (empty) or p42 AUF-1, or with the pENTR/U6 vector carrying a shRNA specific for LacZ or all AUF - 1 mRNAs. Inclusion of APP exons 7 and 8 was assayed by RT-qPCR with primers specific for APP770, APP695 and APP751 . Total APP cDNA was used for normalization. Bars in graphs correspond to mean ± standard deviation of three independent experiments. ( D , E ) SK-N-SH cells were co-transfected with the human ( D ) or the mouse ( E ) APPE78 minigene along with two expression vectors, one bearing ELAVL4 and the other one carrying no insert (empty) or p42 AUF-1, in ratios depicted. Splicing pathways were determined by RT-PCR, as described in Fig. 5 . Quantification of the results was performed by scanning densitometry. Note, that AUF-1 enhances ELAVL4-mediated exclusion of APP exons 7 and 8. ( F ) Neuro2a cells were transfected with the pSilencer vector bearing either no insert (empty) or a shRNA targeting all Auf - 1 transcripts. Inclusion of APP exons 7 and 8 was assayed by RT-qPCR, as described above (**P

    Journal: Scientific Reports

    Article Title: Neuronal ELAVL proteins utilize AUF-1 as a co-partner to induce neuron-specific alternative splicing of APP

    doi: 10.1038/srep44507

    Figure Lengend Snippet: AUF-1 facilitates ELAVL4-mediated APP695-specific AS. Biotinylated RNA probes transcribed from human ( A ) and mouse ( B ) APP minigenes were tested for AUF-1 binding. Note, that AUF-1 isoforms p42 and/or p45 bind to all transcripts shown to interact with nELAVLs. ( C ) SK-N-SH cells were transfected with the DNA3.1 expression vector bearing either no insert (empty) or p42 AUF-1, or with the pENTR/U6 vector carrying a shRNA specific for LacZ or all AUF - 1 mRNAs. Inclusion of APP exons 7 and 8 was assayed by RT-qPCR with primers specific for APP770, APP695 and APP751 . Total APP cDNA was used for normalization. Bars in graphs correspond to mean ± standard deviation of three independent experiments. ( D , E ) SK-N-SH cells were co-transfected with the human ( D ) or the mouse ( E ) APPE78 minigene along with two expression vectors, one bearing ELAVL4 and the other one carrying no insert (empty) or p42 AUF-1, in ratios depicted. Splicing pathways were determined by RT-PCR, as described in Fig. 5 . Quantification of the results was performed by scanning densitometry. Note, that AUF-1 enhances ELAVL4-mediated exclusion of APP exons 7 and 8. ( F ) Neuro2a cells were transfected with the pSilencer vector bearing either no insert (empty) or a shRNA targeting all Auf - 1 transcripts. Inclusion of APP exons 7 and 8 was assayed by RT-qPCR, as described above (**P

    Article Snippet: Quantitative real-time PCR (qPCR) was carried out in the Light Cycler 96 instrument (Roche) using Platinum Taq polymerase (Invitrogen), SYBR Green Reagent (Roche) and primers specific for either each APP isoform cDNA or APP genomic sequences; total APP cDNA and intronic GAPDH were used for normalization (see ).

    Techniques: Binding Assay, Transfection, Expressing, Plasmid Preparation, shRNA, Quantitative RT-PCR, Standard Deviation, Reverse Transcription Polymerase Chain Reaction