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reverse transcriptase enzyme  (Promega)

 
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    Promega reverse transcriptase enzyme
    Reverse Transcriptase Enzyme, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/transcriptase+enzyme/reverse+transcriptase+enzyme/pm37313503-86-23-26
    Average 90 stars, based on 1 article reviews
    reverse transcriptase enzyme - by Bioz Stars, 2026-09
    90/100 stars

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    other:

    Article Title: The intensity of the transcriptional response varies across infection with distinct viral strains in an insect host.
    Article Snippet: Briefly, RNA was isolated from whole bees using an RNeasy mini kit (Qiagen, Hilden, Germany). cDNA was synthesized from 800ng of RNA using oligo(dT)18 primers (Thermo Scientific, Waltham, MA, USA) and reverse transcriptase (M-MLV and Revertase, Promega, Mannheim, Germany) following the manufacturer’s protocol. cDNA was diluted 1:10x prior to qPCR reactions. qPCR was conducted using SYBRgreen Sensimix (Bioline, Luckenwalde, Germany) melt curve analysis to ensure the correct product had been amplified.

    Article Title: Maintaining Toll signaling in Drosophila brain is required to sustain autophagy for dopamine neuron survival
    Article Snippet: Total RNA (2–3 μg) from each sample was used for cDNA synthesis using reverse transcriptase (RTase) (Promega, USA) and oligo dT 18 primer (Intergrated DNA Technologies, USA) at 42°C for 60 min in a 25-μL reaction volume.

    Synthesized:

    Article Title: Mapping Arginase Expression with 18 F-Fluorinated Late-Generation Arginase Inhibitors Derived from Quaternary α-Amino Acids.
    Article Snippet: .. 62 No. 8 August 2021 AppliedBiosystems) according to themanufacturer’s instructions.Complementary DNAwas synthesized from equal amounts of RNAusing the Reverse Transcriptase System (Promega) and the following protocol: 10 min at 25 C, 45 min at 42 C, and 5 min at 99 C. Reverse-transcription polymerase chain reaction was performed with SYBR Green (Roche Diagnostics) and the following protocol, including a final step to generate themelting curve: 2 min at 95 C, 10min at 95 C, 453 (30 s at 95 C, 30 s at 60 C, and 30 s at 72 C), 30 s at 95 C, 30 s at 55 C, and 30 s at 95 C. The reverse-transcription polymerase chain reaction was performed in an Eco system (Illumina). ..

    Reverse Transcription:

    Article Title: Mapping Arginase Expression with 18 F-Fluorinated Late-Generation Arginase Inhibitors Derived from Quaternary α-Amino Acids.
    Article Snippet: .. 62 No. 8 August 2021 AppliedBiosystems) according to themanufacturer’s instructions.Complementary DNAwas synthesized from equal amounts of RNAusing the Reverse Transcriptase System (Promega) and the following protocol: 10 min at 25 C, 45 min at 42 C, and 5 min at 99 C. Reverse-transcription polymerase chain reaction was performed with SYBR Green (Roche Diagnostics) and the following protocol, including a final step to generate themelting curve: 2 min at 95 C, 10min at 95 C, 453 (30 s at 95 C, 30 s at 60 C, and 30 s at 72 C), 30 s at 95 C, 30 s at 55 C, and 30 s at 95 C. The reverse-transcription polymerase chain reaction was performed in an Eco system (Illumina). ..

    Article Title: Sphingomonas Paucimobilis -derived Extracellular Vesicles Reverse Aβ-induced Dysregulation of Neurotrophic Factors, Mitochondrial Function, and Inflammatory Factors through MeCP2-mediated Mechanism.
    Article Snippet: .. Two μg of total RNA was converted to cDNA using a reverse transcriptase system (Promega, Madison, WI, USA). .. The PCR was carried out using the CFX 96 Real-Time PCR System Detector (Bio-Rad Laboratories) in a 20 μl volume containing 4 μl of 1/8 diluted cDNA, 10 μl of 2X iQTM SYBR Green Supermix (Bio-Rad Laboratories, Foster City, CA, USA), and 1 μl each of 5 pmol/μl forward and reverse primers.

    Article Title: The EGR1-Artemin Axis in Keratinocytes Enhances the Innervation of Epidermal Sensory Neurons during Skin Inflammation Induced by House Dust Mite Extract from Dermatophagoidesfarinae.
    Article Snippet: Epidermal hyperinnervation is a critical feature of pruritus during skin inflammation.. However, the mechanisms underlying epidermal hyperinnervation are unclear.. This study investigates the role of the transcription factor EGR1 in epidermal innervation by utilizing wild-type (Egr1þ/þ) and Egr1-null (Egr1) mice topically applied Dermatophagoides farinae extract from dust mite.

    Article Title: Single-Cell Transcriptional Analysis of Lamina Propria Lymphocytes in the Jejunum Reveals Innate Lymphoid Cell-like Cells in Pigs.
    Article Snippet: RESEARCH ARTICLE | NOVEMBER 17 2023 Single-Cell Transcriptional Analysis of Lamina Propria Lymphocytes in the Jejunum Reveals Innate Lymphoid Cell–like Cells in Pigs Junhong Wang; ... et. al J Immunol (2024) 212 (1): 130–142. https://doi.org/10.4049/jimmunol.2300463 Related Content Senescent Jejunal Mast Cells and Eosinophils in the Mouse Preferentially Translocate to the Spleen and Draining Lymph Node, Respectively, During the Recovery Phase of Helminth Infection J Immunol (July,2000) Cathelicidin-WA Improves Intestinal Epithelial Barrier Function and Enhances Host Defense against Enterohemorrhagic Escherichia coli O157:H7 Infection J Immunol (February,2017) D ow nloaded from http://journals.aai.org/jim m unol/article-pdf/212/1/130/1650261/ji2300463.pdf by guest on 03 M ay 2025

    Article Title: Anticancer actions of carnosine in cellular models of prostate cancer.
    Article Snippet: Total RNA was extracted using the RNeasy Mini Extraction Kit (QIAGEN) according to the manufacturer's protocol and the concentration of extracted RNA was measured on a NanoDrop spectrophotometer (Thermo Fisher). .. To synthesize cDNA, 2 μg of RNA in nuclease-free water (final volume 9 μL) and 1 μL of oligo dT (Promega) were heated at 70°C for 5 min. A mix containing 5 μL of RT buffer (Promega), 1 μL of Reverse Transcriptase enzyme (Promega), 0.7 μL of RNasin (Promega), 1 μL of dNTPs (Promega) and 7.3 μL of nuclease-free water were then added to the reaction and incubated at 40°C for 1 h. The reaction was heated at 95°C for 5 min and frozen immediately at −20°C. .. The RT-qPCR reaction was performed in a final volume of 13 μL by mixing 1 μL of cDNA, 6.25 μL of iTaq Universal SYBR Green supermix (Bio-Rad), 0.5 μL of forward and reverse primer (10 pM) and 4.75 μL of nuclease-free water.

    Article Title: Hermetia illucens -Derived Chitosan: A Promising Immunomodulatory Agent for Applications in Biomedical Fields.
    Article Snippet: .. The extracted mRNA was employed to synthesize complementary DNA (cDNA), using reverse transcriptase enzyme (Promega). ..

    Polymerase Chain Reaction:

    Article Title: Mapping Arginase Expression with 18 F-Fluorinated Late-Generation Arginase Inhibitors Derived from Quaternary α-Amino Acids.
    Article Snippet: .. 62 No. 8 August 2021 AppliedBiosystems) according to themanufacturer’s instructions.Complementary DNAwas synthesized from equal amounts of RNAusing the Reverse Transcriptase System (Promega) and the following protocol: 10 min at 25 C, 45 min at 42 C, and 5 min at 99 C. Reverse-transcription polymerase chain reaction was performed with SYBR Green (Roche Diagnostics) and the following protocol, including a final step to generate themelting curve: 2 min at 95 C, 10min at 95 C, 453 (30 s at 95 C, 30 s at 60 C, and 30 s at 72 C), 30 s at 95 C, 30 s at 55 C, and 30 s at 95 C. The reverse-transcription polymerase chain reaction was performed in an Eco system (Illumina). ..

    SYBR Green Assay:

    Article Title: Mapping Arginase Expression with 18 F-Fluorinated Late-Generation Arginase Inhibitors Derived from Quaternary α-Amino Acids.
    Article Snippet: .. 62 No. 8 August 2021 AppliedBiosystems) according to themanufacturer’s instructions.Complementary DNAwas synthesized from equal amounts of RNAusing the Reverse Transcriptase System (Promega) and the following protocol: 10 min at 25 C, 45 min at 42 C, and 5 min at 99 C. Reverse-transcription polymerase chain reaction was performed with SYBR Green (Roche Diagnostics) and the following protocol, including a final step to generate themelting curve: 2 min at 95 C, 10min at 95 C, 453 (30 s at 95 C, 30 s at 60 C, and 30 s at 72 C), 30 s at 95 C, 30 s at 55 C, and 30 s at 95 C. The reverse-transcription polymerase chain reaction was performed in an Eco system (Illumina). ..

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: The EGR1-Artemin Axis in Keratinocytes Enhances the Innervation of Epidermal Sensory Neurons during Skin Inflammation Induced by House Dust Mite Extract from Dermatophagoidesfarinae.
    Article Snippet: Epidermal hyperinnervation is a critical feature of pruritus during skin inflammation.. However, the mechanisms underlying epidermal hyperinnervation are unclear.. This study investigates the role of the transcription factor EGR1 in epidermal innervation by utilizing wild-type (Egr1þ/þ) and Egr1-null (Egr1) mice topically applied Dermatophagoides farinae extract from dust mite.

    Virus:

    Article Title: Single-Cell Transcriptional Analysis of Lamina Propria Lymphocytes in the Jejunum Reveals Innate Lymphoid Cell-like Cells in Pigs.
    Article Snippet: RESEARCH ARTICLE | NOVEMBER 17 2023 Single-Cell Transcriptional Analysis of Lamina Propria Lymphocytes in the Jejunum Reveals Innate Lymphoid Cell–like Cells in Pigs Junhong Wang; ... et. al J Immunol (2024) 212 (1): 130–142. https://doi.org/10.4049/jimmunol.2300463 Related Content Senescent Jejunal Mast Cells and Eosinophils in the Mouse Preferentially Translocate to the Spleen and Draining Lymph Node, Respectively, During the Recovery Phase of Helminth Infection J Immunol (July,2000) Cathelicidin-WA Improves Intestinal Epithelial Barrier Function and Enhances Host Defense against Enterohemorrhagic Escherichia coli O157:H7 Infection J Immunol (February,2017) D ow nloaded from http://journals.aai.org/jim m unol/article-pdf/212/1/130/1650261/ji2300463.pdf by guest on 03 M ay 2025

    Incubation:

    Article Title: Anticancer actions of carnosine in cellular models of prostate cancer.
    Article Snippet: Total RNA was extracted using the RNeasy Mini Extraction Kit (QIAGEN) according to the manufacturer's protocol and the concentration of extracted RNA was measured on a NanoDrop spectrophotometer (Thermo Fisher). .. To synthesize cDNA, 2 μg of RNA in nuclease-free water (final volume 9 μL) and 1 μL of oligo dT (Promega) were heated at 70°C for 5 min. A mix containing 5 μL of RT buffer (Promega), 1 μL of Reverse Transcriptase enzyme (Promega), 0.7 μL of RNasin (Promega), 1 μL of dNTPs (Promega) and 7.3 μL of nuclease-free water were then added to the reaction and incubated at 40°C for 1 h. The reaction was heated at 95°C for 5 min and frozen immediately at −20°C. .. The RT-qPCR reaction was performed in a final volume of 13 μL by mixing 1 μL of cDNA, 6.25 μL of iTaq Universal SYBR Green supermix (Bio-Rad), 0.5 μL of forward and reverse primer (10 pM) and 4.75 μL of nuclease-free water.



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    Image Search Results


    Several RNAs transcribed by the sibD minimal promoter from E. coli chromosomal DNA could be NAD capped. ( A ) Schematic illustration of gene editing designs in the E. coli genome for four small RNAs expression driven by the sibD minimal promoter. The gene body of trpT is highlighted in purple, sroC in green, and ryjA and symR in blue. The sibD minimal promoter ( sibD P-35 ) is labelled as a short red line, and the rrnB terminator is highlighted in yellow. ( B ) Detection of NAD caps in SibD, TrpT, RyjA, SroC, and SymR RNAs with NADbio-northern blotting in the wild-type and indicated mutant strains. ‘ADPRC+’ indicates the biotinylation of NAD-RNAs via the ADPRC-SPAAC reaction with sufficient ADPRC, while ‘ADPRC−’ denotes the ADPRC-SPAAC reaction without ADPRC. 5S RNAs were detected as loading controls. ( C ) Detection and quantification of NAD-RNAs from SibD, TrpT, RyjA, SroC, and SymR using APB gel blotting. Capping ratios were calculated based on the band intensity of the capped transcripts relative to the total transcripts (both capped and uncapped transcripts) in the APB gel.

    Journal: Nucleic Acids Research

    Article Title: NAD + capping of sibD transcripts in E. coli is mediated by its minimal promoter and enhanced by ppGpp

    doi: 10.1093/nar/gkag102

    Figure Lengend Snippet: Several RNAs transcribed by the sibD minimal promoter from E. coli chromosomal DNA could be NAD capped. ( A ) Schematic illustration of gene editing designs in the E. coli genome for four small RNAs expression driven by the sibD minimal promoter. The gene body of trpT is highlighted in purple, sroC in green, and ryjA and symR in blue. The sibD minimal promoter ( sibD P-35 ) is labelled as a short red line, and the rrnB terminator is highlighted in yellow. ( B ) Detection of NAD caps in SibD, TrpT, RyjA, SroC, and SymR RNAs with NADbio-northern blotting in the wild-type and indicated mutant strains. ‘ADPRC+’ indicates the biotinylation of NAD-RNAs via the ADPRC-SPAAC reaction with sufficient ADPRC, while ‘ADPRC−’ denotes the ADPRC-SPAAC reaction without ADPRC. 5S RNAs were detected as loading controls. ( C ) Detection and quantification of NAD-RNAs from SibD, TrpT, RyjA, SroC, and SymR using APB gel blotting. Capping ratios were calculated based on the band intensity of the capped transcripts relative to the total transcripts (both capped and uncapped transcripts) in the APB gel.

    Article Snippet: To perform IVT assays with various sigma factors, a similar assay was conducted, except the E. coli RNAP holoenzyme was replaced by the same amount of E. coli RNAP core enzyme (NEB) and in the absence of ppGpp or DksA.

    Techniques: Expressing, Northern Blot, Mutagenesis

    Effects of (p)ppGpp and DksA on transcription and NAD capping of certain small RNAs in E. coli cells. ( A ) The NAD capping level of SibD increased upon transient induction of RelA 455aa and DksA. Both RelA 455aa and DksA were expressed from plasmids under the control of the pBAD promoter. NAD capping of SibD was assessed by APB gel blotting. The total level of SibD RNA in each lane was quantified from the normal gel using ImageJ software and normalized to the intensity in the first EV lane. The NAD capping ratio was calculated as the percentage of the intensity of the NAD-capped band relative to the sum of the intensities of both the capped and uncapped bands in the APB gel. ‘Arabinose−’ indicates RNA samples without arabinose induction, while ‘Arabinose+’ signifies that arabinose was added to induce the expression of RelA 455aa and DksA. ‘EV’ indicates strain carrying the empty pBAD33.1 vector. The tmRNA was used as a loading control and each blotting has three independent replicates. ( B ) Detection of NAD-capped transcripts of five sRNAs with NADbio-northern blotting analysis, including four known NAD-RNAs: SibC, SibD, SibE, and GcvB. The tmRNA was used as a loading control. ( C – G ) Detection of total transcripts and NAD-capped transcripts of five sRNAs, namely SibA, SibC, SibD, SibE, and GcvB, respectively. The total abundance of individual RNA was determined by electrophoresis on a standard PAGE gel followed by northern blotting (labelled as normal gel), while the NAD-capped transcripts were identified with APB gel blotting (labelled as APB gel). The non-NAD-RNA SibA was included as a negative control. The NAD capping ratio was calculated based on the band intensity of the NAD-capped version relative to the total transcription levels (NAD-capped version plus uncapped version). Two types of synthetic RNAs for each sRNA, namely with 5′-ppp- and 5′-NAD modifications, were used as controls.

    Journal: Nucleic Acids Research

    Article Title: NAD + capping of sibD transcripts in E. coli is mediated by its minimal promoter and enhanced by ppGpp

    doi: 10.1093/nar/gkag102

    Figure Lengend Snippet: Effects of (p)ppGpp and DksA on transcription and NAD capping of certain small RNAs in E. coli cells. ( A ) The NAD capping level of SibD increased upon transient induction of RelA 455aa and DksA. Both RelA 455aa and DksA were expressed from plasmids under the control of the pBAD promoter. NAD capping of SibD was assessed by APB gel blotting. The total level of SibD RNA in each lane was quantified from the normal gel using ImageJ software and normalized to the intensity in the first EV lane. The NAD capping ratio was calculated as the percentage of the intensity of the NAD-capped band relative to the sum of the intensities of both the capped and uncapped bands in the APB gel. ‘Arabinose−’ indicates RNA samples without arabinose induction, while ‘Arabinose+’ signifies that arabinose was added to induce the expression of RelA 455aa and DksA. ‘EV’ indicates strain carrying the empty pBAD33.1 vector. The tmRNA was used as a loading control and each blotting has three independent replicates. ( B ) Detection of NAD-capped transcripts of five sRNAs with NADbio-northern blotting analysis, including four known NAD-RNAs: SibC, SibD, SibE, and GcvB. The tmRNA was used as a loading control. ( C – G ) Detection of total transcripts and NAD-capped transcripts of five sRNAs, namely SibA, SibC, SibD, SibE, and GcvB, respectively. The total abundance of individual RNA was determined by electrophoresis on a standard PAGE gel followed by northern blotting (labelled as normal gel), while the NAD-capped transcripts were identified with APB gel blotting (labelled as APB gel). The non-NAD-RNA SibA was included as a negative control. The NAD capping ratio was calculated based on the band intensity of the NAD-capped version relative to the total transcription levels (NAD-capped version plus uncapped version). Two types of synthetic RNAs for each sRNA, namely with 5′-ppp- and 5′-NAD modifications, were used as controls.

    Article Snippet: To perform IVT assays with various sigma factors, a similar assay was conducted, except the E. coli RNAP holoenzyme was replaced by the same amount of E. coli RNAP core enzyme (NEB) and in the absence of ppGpp or DksA.

    Techniques: Control, Software, Expressing, Plasmid Preparation, Northern Blot, Electrophoresis, Negative Control