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Promega amv reverse transcriptase
Amv Reverse Transcriptase, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 229 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 229 article reviews
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amv reverse transcriptase - by Bioz Stars, 2020-02
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Related Articles

Clone Assay:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: Paragraph title: Mutation mapping and cloning: ... Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI).

Amplification:

Article Title: Molecular Identification of a Novel Hantavirus in Malaysian Bronze Tube-Nosed Bats (Murina aenea)
Article Snippet: .. Samples were then subjected to the Sequence Independent Single Primer Amplification (SISPA) approach [ ]. cDNA amplification was performed by an AMV Reverse Transcriptase (Promega) according to the provided manual by the manufacturers using the FR26RV-N (5′-GCCGGAGCTCTGCAGATATCNNNNNN-3′) primer. .. Thereafter, ds cDNA was amplified by a DreamTaq DNA Polymerase (Thermo Fisher Scientific, Waltham, MA, USA) according to the supplied protocol using the FR20RV (5′-GCCGGAGCTCTGCAGATATC-3′) primer.

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: .. Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific). .. Resulting DNA smears extracted from gel slices were used for library preparation compatible with semiconductor sequencing.

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: The tail DNA samples of N2 mice were extracted and PCR amplified with D5Mit356 and D5Mit359 microsatellite markers of the Whitehead/MIT database ( ). .. Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI).

Article Title: Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia
Article Snippet: Then, RNA was reverse-transcribed into cDNA according to the instructions of the reverse transcription kit, followed by PCR amplification based on the amplification system (a total of 35 cycles). .. In brief, after determining the RNA concentration, in vitro reverse transcription was performed in a volume of 20 μL with a system including 4 μl 25 mM MgCl2 , 2 μL reverse transcription 10×buffer, 2 μL 104 mM dNTP mixture, 0.5 μL ribonuclease inhibitor, 15U AMV reverse transcriptase, 0.5 μg random primers, 1μ total RNA and nuclease-free water to a final volume of 20 μL at 42°C for 15 min and 85°C denaturing (Promega, USA).

Synthesized:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: .. Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI). .. Due to the large size of c-kit cDNA (2925 bp), three overlapping cDNA fragments that covered the full length were then amplified by PCR with LA-Taq polymerase (TaKaRa, Shiga, Japan) using the following primers: kit1-FOR, 5′-TCAGAGTCTAGCGCAGCCAC-3′; kit1-REV, 5′-GCCTCGTATTCAACAACCAA-3′; kit2-FOR, 5′-TGTAACCGATGGAGAAAACG-3′; kit2-REV, 5′-TAAACGAGTCACGCTTCCTT-3′; kit3-FOR, 5′-GCCCTAATGTCGGAACTGAA-3′; and kit3-REV, 5′-GTTTCTGCTCAGGCATCTTC-3′.

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: .. Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA). .. The primer sequences used were as follows: human PTK2 (FAK, NCBI reference sequence: NM_001199649.1), forward: 5′-TTA TTG GCC ACT GTG GAT GA-3′, reverse: 5′-TAC TCT TGC TGG AGG CTG GT-3′; human TLN1 (Talin1, NCBI reference sequence: NM_006289.3), forward: 5′-TCT CCC AAA ATG CCA AGA AC-3′, reverse: 5′-CTC CAC TAG CCC TTG CTG TC-3′; human VCL (vinculin, NCBI reference sequence: NM_003373.3), forward: 5′-GCC AAG CAG TGC ACA GAT AA-3′, reverse: 5′-AGG TTC TGG GCA TTG TGA AC-3′, human PXN (paxillin, NCBI reference sequence: NM_001080855.2), forward: 5′-GGA GTC TAC CAC CTC CCA CA-3′, reverse: 5′-CCA CTG GTC TAA GGG GTC AA-3′, human GAPDH (NCBI reference sequence: NM_001256799.2), forward: 5′-CAT GGG TGT GAA CCA TGA GAA-3′, reverse: 5′-GGT CAT GAG TCC TTC CAC GAT-3′.

Article Title: Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome
Article Snippet: .. Single-stranded cDNA was synthesized with AMV reverse transcriptase and random hexamers (Promega). ..

TA Cloning:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI). .. The three fragments were then subcloned into TA cloning vector pMD 18-T (TaKaRa) and sequenced (Bioasia, Shanghai, China).

Quantitative RT-PCR:

Article Title: LncRNA TP73-AS1 Activates TGF-β1 to Promote the Migration and Invasion of Colorectal Cancer Cell
Article Snippet: Paragraph title: Total RNA Extraction and RT-qPCR ... Following cDNA synthesis using AMV Reverse Transcriptase (Promega Corporation, USA), qPCR reaction systems were prepared using SYBR® Green master mix (Bio-Rad, USA) with 18S rRNA or GAPDH as endogenous controls to detect the expression of TP73-AS1 and TGF-β1, respectively.

Article Title: Reduction of the vitamin D hormonal system in kidney disease is associated with increased renal inflammation
Article Snippet: Paragraph title: Quantitative (real-time) RT-PCR analysis of CYP27B1, CYP24A1 VDR, and MCP-1 mRNA in renal biopsies ... Aliquots (1.5 µg) of RNA from each biopsy were then reverse transcribed using AMV reverse transcriptase (Promega, Southampton, UK).

Article Title: lncRNA DGCR5 Up-Regulates TGF-β1, Increases Cancer Cell Stemness and Predicts Survival of Prostate Cancer Patients
Article Snippet: .. RT-qPCR RNAzol reagent (Sigma-Aldrich, USA) was used to extract total RNAs from PC tissues, non-cancer tissues as well as cells of both 22Rv1 and DU145 cell lines to perform reverse transcription, which was carried out using AMV Reverse Transcriptase (Promega Corporation). .. To detect the expression of DGCR5 and TGF-β1, PCR systems were prepared using Applied Biosystems™ PowerUp™ SYBR™ Green Master Mix (Thermo Fisher Scientific, Inc.) with 18S rRNA as the endogenous control of DGCR5 and GAPDH as the endogenous control of TGF-β1.

SYBR Green Assay:

Article Title: LncRNA TP73-AS1 Activates TGF-β1 to Promote the Migration and Invasion of Colorectal Cancer Cell
Article Snippet: .. Following cDNA synthesis using AMV Reverse Transcriptase (Promega Corporation, USA), qPCR reaction systems were prepared using SYBR® Green master mix (Bio-Rad, USA) with 18S rRNA or GAPDH as endogenous controls to detect the expression of TP73-AS1 and TGF-β1, respectively. .. PCR reactions were repeated 3 times.

Article Title: lncRNA DGCR5 Up-Regulates TGF-β1, Increases Cancer Cell Stemness and Predicts Survival of Prostate Cancer Patients
Article Snippet: RT-qPCR RNAzol reagent (Sigma-Aldrich, USA) was used to extract total RNAs from PC tissues, non-cancer tissues as well as cells of both 22Rv1 and DU145 cell lines to perform reverse transcription, which was carried out using AMV Reverse Transcriptase (Promega Corporation). .. To detect the expression of DGCR5 and TGF-β1, PCR systems were prepared using Applied Biosystems™ PowerUp™ SYBR™ Green Master Mix (Thermo Fisher Scientific, Inc.) with 18S rRNA as the endogenous control of DGCR5 and GAPDH as the endogenous control of TGF-β1.

Random Hexamer Labeling:

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: .. Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific). .. Resulting DNA smears extracted from gel slices were used for library preparation compatible with semiconductor sequencing.

Expressing:

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Because Q6NUR6 gene is close to, but not interrupted by, the 7p22.1 breakpoint, we wanted to determine if its expression is altered by the presence of the translocation breakpoint. .. Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA.

Article Title: LncRNA TP73-AS1 Activates TGF-β1 to Promote the Migration and Invasion of Colorectal Cancer Cell
Article Snippet: .. Following cDNA synthesis using AMV Reverse Transcriptase (Promega Corporation, USA), qPCR reaction systems were prepared using SYBR® Green master mix (Bio-Rad, USA) with 18S rRNA or GAPDH as endogenous controls to detect the expression of TP73-AS1 and TGF-β1, respectively. .. PCR reactions were repeated 3 times.

Article Title: N-Palmitoylethanolamide-Oxazoline Protects against Middle Cerebral Artery Occlusion Injury in Diabetic Rats by Regulating the SIRT1 Pathway
Article Snippet: RT-PCR Total RNA was extracted according to the manufacturer’s protocol and reverse-transcribed using 2-μg oligo(dT)15 primer, 10 units of AMV reverse transcriptase, 40 units of RNase inhibitor (all from Promega, Southampton, UK), and 1.25 mM of dNTP (Bioline, London, UK) for 45 min at 42 °C. .. Sequence-specific fluorescent signals were detected by an ABI Prism 7700Sequence Detector System, and mRNA data were normalized relative to GADPH and then used to calculate expression levels.

Article Title: Hypoxia enhances CXCR4 expression in human microvascular endothelial cells and human melanoma cells
Article Snippet: In order to investigate the influence of matrix components, serum factors and oxygen concentrations on chemokine receptor mRNA expression, HMEC-1 cells (1×106 cells/p60 plate) were plated on various basement membrane matrix components, namely gelatin (0.1%), collagen type IV (20 μg/ml) or Matrigel (0.25 mg/ml). .. Next, 1μg of RNA was reverse transcribed using AMV reverse transcriptase (AMV-RT; 0.2 U/μl; Promega, Madison, WI, USA) and its buffer in the presence of oligod(T)16 primers (1 μM; Applied Biosystems, Foster City, CA, USA), 0.2 mM of each of the dNTPs (Sigma-Aldrich) and RNAse inhibitor (RNAsin, 0.4 U/μl; Promega) in a 25 μl reaction mixture.

Article Title: lncRNA DGCR5 Up-Regulates TGF-β1, Increases Cancer Cell Stemness and Predicts Survival of Prostate Cancer Patients
Article Snippet: RT-qPCR RNAzol reagent (Sigma-Aldrich, USA) was used to extract total RNAs from PC tissues, non-cancer tissues as well as cells of both 22Rv1 and DU145 cell lines to perform reverse transcription, which was carried out using AMV Reverse Transcriptase (Promega Corporation). .. To detect the expression of DGCR5 and TGF-β1, PCR systems were prepared using Applied Biosystems™ PowerUp™ SYBR™ Green Master Mix (Thermo Fisher Scientific, Inc.) with 18S rRNA as the endogenous control of DGCR5 and GAPDH as the endogenous control of TGF-β1.

Real-time Polymerase Chain Reaction:

Article Title: Effect of Dietary Nutrient Density on Small Intestinal Phosphate Transport and Bone Mineralization of Broilers during the Growing Period
Article Snippet: Paragraph title: Total RNA extraction, reverse transcription, and real-time PCR ... Afterward, 1.0 μg of total RNA was reverse-transcribed into single-stranded cDNA with AMV reverse transcriptase and an Oligo (dT)15 primer in the presence of recombinant RNasin ribonuclease inhibitor (A3500; Promega Corporation).

Article Title: LncRNA TP73-AS1 Activates TGF-β1 to Promote the Migration and Invasion of Colorectal Cancer Cell
Article Snippet: .. Following cDNA synthesis using AMV Reverse Transcriptase (Promega Corporation, USA), qPCR reaction systems were prepared using SYBR® Green master mix (Bio-Rad, USA) with 18S rRNA or GAPDH as endogenous controls to detect the expression of TP73-AS1 and TGF-β1, respectively. .. PCR reactions were repeated 3 times.

Article Title: N-Palmitoylethanolamide-Oxazoline Protects against Middle Cerebral Artery Occlusion Injury in Diabetic Rats by Regulating the SIRT1 Pathway
Article Snippet: RT-PCR Total RNA was extracted according to the manufacturer’s protocol and reverse-transcribed using 2-μg oligo(dT)15 primer, 10 units of AMV reverse transcriptase, 40 units of RNase inhibitor (all from Promega, Southampton, UK), and 1.25 mM of dNTP (Bioline, London, UK) for 45 min at 42 °C. .. Real-time PCR was carried out using TaqMan Universal PCR master mix and fluorescent primers obtained from Quiagen (QuantiTect primers, Skelton House, UK).

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: Paragraph title: Total RNA extraction, reverse transcription, and real-time PCR ... After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.).

Translocation Assay:

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Because Q6NUR6 gene is close to, but not interrupted by, the 7p22.1 breakpoint, we wanted to determine if its expression is altered by the presence of the translocation breakpoint. .. Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA.

Countercurrent Chromatography:

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA). .. The primer sequences used were as follows: human PTK2 (FAK, NCBI reference sequence: NM_001199649.1), forward: 5′-TTA TTG GCC ACT GTG GAT GA-3′, reverse: 5′-TAC TCT TGC TGG AGG CTG GT-3′; human TLN1 (Talin1, NCBI reference sequence: NM_006289.3), forward: 5′-TCT CCC AAA ATG CCA AGA AC-3′, reverse: 5′-CTC CAC TAG CCC TTG CTG TC-3′; human VCL (vinculin, NCBI reference sequence: NM_003373.3), forward: 5′-GCC AAG CAG TGC ACA GAT AA-3′, reverse: 5′-AGG TTC TGG GCA TTG TGA AC-3′, human PXN (paxillin, NCBI reference sequence: NM_001080855.2), forward: 5′-GGA GTC TAC CAC CTC CCA CA-3′, reverse: 5′-CCA CTG GTC TAA GGG GTC AA-3′, human GAPDH (NCBI reference sequence: NM_001256799.2), forward: 5′-CAT GGG TGT GAA CCA TGA GAA-3′, reverse: 5′-GGT CAT GAG TCC TTC CAC GAT-3′.

Southern Blot:

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: To test whether this part of the gene was interrupted by the translocation breakpoint we performed Southern blot experiments to explore the genomic region containing the Q6NUR6 gene and its promoter on the patient's and a control DNA. .. Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA.

Cell Culture:

Article Title: Dermcidin expression in hepatic cells improves survival without N-glycosylation, but requires asparagine residues
Article Snippet: RNA preparation and reverse transcription RNA was prepared from cells cultured in six-well plates using Trizol (Life Technologies, Paisley, UK) according to the manufacturer's instructions. .. Reverse transcription was performed in 20 μ l reactions containing 10 μ l of RNA sample plus 10 μ l of RT mix (4 μ l 25 mM MgCl2 , 2 μ l reverse transcription 10 × buffer, 2 μ l dNTP mixture, 1 μ l oligo(dT)15 primer, 0.5 μ l AMV reverse transcriptase and 0.5 μ l recombinant RNasin ribonuclease inhibitor, all from Promega).

Generated:

Article Title: Hypoxia enhances CXCR4 expression in human microvascular endothelial cells and human melanoma cells
Article Snippet: Next, 1μg of RNA was reverse transcribed using AMV reverse transcriptase (AMV-RT; 0.2 U/μl; Promega, Madison, WI, USA) and its buffer in the presence of oligod(T)16 primers (1 μM; Applied Biosystems, Foster City, CA, USA), 0.2 mM of each of the dNTPs (Sigma-Aldrich) and RNAse inhibitor (RNAsin, 0.4 U/μl; Promega) in a 25 μl reaction mixture. .. In parallel, reactions were carried out in the absence of reverse transcriptase as negative controls (non-RT), whereas cDNA generated from PBMC and MCF7 cells could serve as positive controls.

Polymerase Chain Reaction:

Article Title: Molecular Identification of a Novel Hantavirus in Malaysian Bronze Tube-Nosed Bats (Murina aenea)
Article Snippet: Samples were then subjected to the Sequence Independent Single Primer Amplification (SISPA) approach [ ]. cDNA amplification was performed by an AMV Reverse Transcriptase (Promega) according to the provided manual by the manufacturers using the FR26RV-N (5′-GCCGGAGCTCTGCAGATATCNNNNNN-3′) primer. .. Amplified cDNA was purified by NucleoSpin® Gel and a PCR Clean-up kit (Macherey-Nagel, Düren, Germany), and quantified using a Qubit dsDNA BR Assay kit (Thermo Fisher Scientific).

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: PCR products were separated by electrophoresis with 4% agarose II (BioBasic, Markham, ON, Canada) gel. .. Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI).

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA. .. For the PCR step, 1 μ g of total RNA converted to cDNA was used in 25 μ L reactions with TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA, USA).

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: The cells were collected at 4 h after the initial seeding for reverse transcriptase PCR. .. Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA).

Article Title: LncRNA TP73-AS1 Activates TGF-β1 to Promote the Migration and Invasion of Colorectal Cancer Cell
Article Snippet: Following cDNA synthesis using AMV Reverse Transcriptase (Promega Corporation, USA), qPCR reaction systems were prepared using SYBR® Green master mix (Bio-Rad, USA) with 18S rRNA or GAPDH as endogenous controls to detect the expression of TP73-AS1 and TGF-β1, respectively. .. PCR reactions were repeated 3 times.

Article Title: N-Palmitoylethanolamide-Oxazoline Protects against Middle Cerebral Artery Occlusion Injury in Diabetic Rats by Regulating the SIRT1 Pathway
Article Snippet: RT-PCR Total RNA was extracted according to the manufacturer’s protocol and reverse-transcribed using 2-μg oligo(dT)15 primer, 10 units of AMV reverse transcriptase, 40 units of RNase inhibitor (all from Promega, Southampton, UK), and 1.25 mM of dNTP (Bioline, London, UK) for 45 min at 42 °C. .. Real-time PCR was carried out using TaqMan Universal PCR master mix and fluorescent primers obtained from Quiagen (QuantiTect primers, Skelton House, UK).

Article Title: Reduction of the vitamin D hormonal system in kidney disease is associated with increased renal inflammation
Article Snippet: Aliquots (1.5 µg) of RNA from each biopsy were then reverse transcribed using AMV reverse transcriptase (Promega, Southampton, UK). .. Gene-specific PCR was then carried out using an ABI 7700 sequence detection system (PE Biosystems, Warring, UK) as described previously, using primers and probes outlined below under the following conditions: 50 °C for 2 min; 95 °C for 10 min; followed by 44 cycles of 95 °C for 15 s and 60 °C for 1 min.

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.). .. The primers used and PCR product lengths are depicted in .

Article Title: Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia
Article Snippet: Then, RNA was reverse-transcribed into cDNA according to the instructions of the reverse transcription kit, followed by PCR amplification based on the amplification system (a total of 35 cycles). .. In brief, after determining the RNA concentration, in vitro reverse transcription was performed in a volume of 20 μL with a system including 4 μl 25 mM MgCl2 , 2 μL reverse transcription 10×buffer, 2 μL 104 mM dNTP mixture, 0.5 μL ribonuclease inhibitor, 15U AMV reverse transcriptase, 0.5 μg random primers, 1μ total RNA and nuclease-free water to a final volume of 20 μL at 42°C for 15 min and 85°C denaturing (Promega, USA).

Article Title: Hypoxia enhances CXCR4 expression in human microvascular endothelial cells and human melanoma cells
Article Snippet: Next, 1μg of RNA was reverse transcribed using AMV reverse transcriptase (AMV-RT; 0.2 U/μl; Promega, Madison, WI, USA) and its buffer in the presence of oligod(T)16 primers (1 μM; Applied Biosystems, Foster City, CA, USA), 0.2 mM of each of the dNTPs (Sigma-Aldrich) and RNAse inhibitor (RNAsin, 0.4 U/μl; Promega) in a 25 μl reaction mixture. .. The PCR reaction mixture (25 μl) contained 1 μl of reverse transcription mixture in the presence of Taq DNA polymerase (0.06 U/μl; Sigma-Aldrich) and its buffer, 0.2 mM of each of the dNTPs and 0.4 μM of the primer pairs.

Article Title: lncRNA DGCR5 Up-Regulates TGF-β1, Increases Cancer Cell Stemness and Predicts Survival of Prostate Cancer Patients
Article Snippet: RT-qPCR RNAzol reagent (Sigma-Aldrich, USA) was used to extract total RNAs from PC tissues, non-cancer tissues as well as cells of both 22Rv1 and DU145 cell lines to perform reverse transcription, which was carried out using AMV Reverse Transcriptase (Promega Corporation). .. To detect the expression of DGCR5 and TGF-β1, PCR systems were prepared using Applied Biosystems™ PowerUp™ SYBR™ Green Master Mix (Thermo Fisher Scientific, Inc.) with 18S rRNA as the endogenous control of DGCR5 and GAPDH as the endogenous control of TGF-β1.

Random Primed:

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: .. Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific). .. Resulting DNA smears extracted from gel slices were used for library preparation compatible with semiconductor sequencing.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: .. Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific). .. Resulting DNA smears extracted from gel slices were used for library preparation compatible with semiconductor sequencing.

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Paragraph title: Reverse transcription polymerase chain reaction (RT-PCR) ... Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA).

Article Title: Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome
Article Snippet: Paragraph title: RT-PCR. ... Single-stranded cDNA was synthesized with AMV reverse transcriptase and random hexamers (Promega).

Article Title: N-Palmitoylethanolamide-Oxazoline Protects against Middle Cerebral Artery Occlusion Injury in Diabetic Rats by Regulating the SIRT1 Pathway
Article Snippet: .. RT-PCR Total RNA was extracted according to the manufacturer’s protocol and reverse-transcribed using 2-μg oligo(dT)15 primer, 10 units of AMV reverse transcriptase, 40 units of RNase inhibitor (all from Promega, Southampton, UK), and 1.25 mM of dNTP (Bioline, London, UK) for 45 min at 42 °C. .. Real-time PCR was carried out using TaqMan Universal PCR master mix and fluorescent primers obtained from Quiagen (QuantiTect primers, Skelton House, UK).

Article Title: Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia
Article Snippet: Paragraph title: Detection of VEGF and sFlt-1 mRNA levels via RT-PCR ... In brief, after determining the RNA concentration, in vitro reverse transcription was performed in a volume of 20 μL with a system including 4 μl 25 mM MgCl2 , 2 μL reverse transcription 10×buffer, 2 μL 104 mM dNTP mixture, 0.5 μL ribonuclease inhibitor, 15U AMV reverse transcriptase, 0.5 μg random primers, 1μ total RNA and nuclease-free water to a final volume of 20 μL at 42°C for 15 min and 85°C denaturing (Promega, USA).

Recombinant:

Article Title: Effect of Dietary Nutrient Density on Small Intestinal Phosphate Transport and Bone Mineralization of Broilers during the Growing Period
Article Snippet: .. Afterward, 1.0 μg of total RNA was reverse-transcribed into single-stranded cDNA with AMV reverse transcriptase and an Oligo (dT)15 primer in the presence of recombinant RNasin ribonuclease inhibitor (A3500; Promega Corporation). .. The mRNAs of NaPi-IIb and NaPi-IIa were subjected to real-time PCR with β-actin as an internal control standard.

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: .. After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.). .. Real-time PCR of NaPi-IIb and calbindin mRNA was performed with β-actin as internal control standard.

Article Title: Dermcidin expression in hepatic cells improves survival without N-glycosylation, but requires asparagine residues
Article Snippet: .. Reverse transcription was performed in 20 μ l reactions containing 10 μ l of RNA sample plus 10 μ l of RT mix (4 μ l 25 mM MgCl2 , 2 μ l reverse transcription 10 × buffer, 2 μ l dNTP mixture, 1 μ l oligo(dT)15 primer, 0.5 μ l AMV reverse transcriptase and 0.5 μ l recombinant RNasin ribonuclease inhibitor, all from Promega). .. Samples were then incubated in a thermal cycler (PHC-3, Techne) for 1 h at 42°C, followed by 5 min at 99°C.

Fluorescence:

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA. .. Fluorescence detection was measured utilizing an ABI Prism 7500 platform (Applied Biosystems).

Mutagenesis:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: Paragraph title: Mutation mapping and cloning: ... Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI).

Isolation:

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: LEDV strains 6101 and 6118 were isolated in 1972 from a pool of 200 C. modestus mosquitoes each. .. Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific).

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Total RNA was isolated and purified using the RNAgents Total RNA Isolation System (Promega) from peripheral white blood cells and EBV cell lines of the patient and controls. .. Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA.

Article Title: Effect of Dietary Nutrient Density on Small Intestinal Phosphate Transport and Bone Mineralization of Broilers during the Growing Period
Article Snippet: Total RNA extraction, reverse transcription, and real-time PCR Total RNA was extracted from the duodenal mucosa and kidney of 42-day-old broilers using the SV Total RNA Isolation System (Z3100; Promega Corporation, Madison, WI, USA) in accordance with the manufacturer’s instructions. .. Afterward, 1.0 μg of total RNA was reverse-transcribed into single-stranded cDNA with AMV reverse transcriptase and an Oligo (dT)15 primer in the presence of recombinant RNasin ribonuclease inhibitor (A3500; Promega Corporation).

Article Title: Reduction of the vitamin D hormonal system in kidney disease is associated with increased renal inflammation
Article Snippet: Total RNA was isolated from frozen kidney biopsies using the GenElute RNA extraction system (Sigma, Poole, UK). .. Aliquots (1.5 µg) of RNA from each biopsy were then reverse transcribed using AMV reverse transcriptase (Promega, Southampton, UK).

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: Total RNA extraction, reverse transcription, and real-time PCR Total RNA was extracted from the duodenal mucosa of 21 d-old broilers with Promega Corporation products according to SV Total RNA Isolation System instructions (Z3100, Promega Corp.), and re-suspended in diethylpyrocarbonate-treated water. .. After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.).

Article Title: Hypoxia enhances CXCR4 expression in human microvascular endothelial cells and human melanoma cells
Article Snippet: After the appropriate incubation of the endothelial and SK-MEL-5 cells, RNA was isolated with the RNeasy kit (Qiagen, Valencia, CA, USA) with an on-column DNAse treatment (RNAse free DNAse set; Qiagen). .. Next, 1μg of RNA was reverse transcribed using AMV reverse transcriptase (AMV-RT; 0.2 U/μl; Promega, Madison, WI, USA) and its buffer in the presence of oligod(T)16 primers (1 μM; Applied Biosystems, Foster City, CA, USA), 0.2 mM of each of the dNTPs (Sigma-Aldrich) and RNAse inhibitor (RNAsin, 0.4 U/μl; Promega) in a 25 μl reaction mixture.

Purification:

Article Title: Molecular Identification of a Novel Hantavirus in Malaysian Bronze Tube-Nosed Bats (Murina aenea)
Article Snippet: Samples were then subjected to the Sequence Independent Single Primer Amplification (SISPA) approach [ ]. cDNA amplification was performed by an AMV Reverse Transcriptase (Promega) according to the provided manual by the manufacturers using the FR26RV-N (5′-GCCGGAGCTCTGCAGATATCNNNNNN-3′) primer. .. Amplified cDNA was purified by NucleoSpin® Gel and a PCR Clean-up kit (Macherey-Nagel, Düren, Germany), and quantified using a Qubit dsDNA BR Assay kit (Thermo Fisher Scientific).

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Total RNA was isolated and purified using the RNAgents Total RNA Isolation System (Promega) from peripheral white blood cells and EBV cell lines of the patient and controls. .. Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA.

Sequencing:

Article Title: Molecular Identification of a Novel Hantavirus in Malaysian Bronze Tube-Nosed Bats (Murina aenea)
Article Snippet: .. Samples were then subjected to the Sequence Independent Single Primer Amplification (SISPA) approach [ ]. cDNA amplification was performed by an AMV Reverse Transcriptase (Promega) according to the provided manual by the manufacturers using the FR26RV-N (5′-GCCGGAGCTCTGCAGATATCNNNNNN-3′) primer. .. Thereafter, ds cDNA was amplified by a DreamTaq DNA Polymerase (Thermo Fisher Scientific, Waltham, MA, USA) according to the supplied protocol using the FR20RV (5′-GCCGGAGCTCTGCAGATATC-3′) primer.

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: LEDV strains were selected for shotgun deep sequencing from cell lysates using the protocols described elsewhere [ ]. .. Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific).

Article Title: Exploring the 7p22.1 Chromosome as a Candidate Region for Autism
Article Snippet: Total RNA was used in conjunction with AMV reverse transcriptase and random hexamers (Promega) to generate first strand cDNA. .. For the Q6NUR6 gene the corresponding sequence-specific primers/probes assay mix (Hs00415445_m1) was used.

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA). .. The primer sequences used were as follows: human PTK2 (FAK, NCBI reference sequence: NM_001199649.1), forward: 5′-TTA TTG GCC ACT GTG GAT GA-3′, reverse: 5′-TAC TCT TGC TGG AGG CTG GT-3′; human TLN1 (Talin1, NCBI reference sequence: NM_006289.3), forward: 5′-TCT CCC AAA ATG CCA AGA AC-3′, reverse: 5′-CTC CAC TAG CCC TTG CTG TC-3′; human VCL (vinculin, NCBI reference sequence: NM_003373.3), forward: 5′-GCC AAG CAG TGC ACA GAT AA-3′, reverse: 5′-AGG TTC TGG GCA TTG TGA AC-3′, human PXN (paxillin, NCBI reference sequence: NM_001080855.2), forward: 5′-GGA GTC TAC CAC CTC CCA CA-3′, reverse: 5′-CCA CTG GTC TAA GGG GTC AA-3′, human GAPDH (NCBI reference sequence: NM_001256799.2), forward: 5′-CAT GGG TGT GAA CCA TGA GAA-3′, reverse: 5′-GGT CAT GAG TCC TTC CAC GAT-3′.

Article Title: N-Palmitoylethanolamide-Oxazoline Protects against Middle Cerebral Artery Occlusion Injury in Diabetic Rats by Regulating the SIRT1 Pathway
Article Snippet: RT-PCR Total RNA was extracted according to the manufacturer’s protocol and reverse-transcribed using 2-μg oligo(dT)15 primer, 10 units of AMV reverse transcriptase, 40 units of RNase inhibitor (all from Promega, Southampton, UK), and 1.25 mM of dNTP (Bioline, London, UK) for 45 min at 42 °C. .. Sequence-specific fluorescent signals were detected by an ABI Prism 7700Sequence Detector System, and mRNA data were normalized relative to GADPH and then used to calculate expression levels.

Article Title: Reduction of the vitamin D hormonal system in kidney disease is associated with increased renal inflammation
Article Snippet: Aliquots (1.5 µg) of RNA from each biopsy were then reverse transcribed using AMV reverse transcriptase (Promega, Southampton, UK). .. Gene-specific PCR was then carried out using an ABI 7700 sequence detection system (PE Biosystems, Warring, UK) as described previously, using primers and probes outlined below under the following conditions: 50 °C for 2 min; 95 °C for 10 min; followed by 44 cycles of 95 °C for 15 s and 60 °C for 1 min.

Activated Clotting Time Assay:

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA). .. The primer sequences used were as follows: human PTK2 (FAK, NCBI reference sequence: NM_001199649.1), forward: 5′-TTA TTG GCC ACT GTG GAT GA-3′, reverse: 5′-TAC TCT TGC TGG AGG CTG GT-3′; human TLN1 (Talin1, NCBI reference sequence: NM_006289.3), forward: 5′-TCT CCC AAA ATG CCA AGA AC-3′, reverse: 5′-CTC CAC TAG CCC TTG CTG TC-3′; human VCL (vinculin, NCBI reference sequence: NM_003373.3), forward: 5′-GCC AAG CAG TGC ACA GAT AA-3′, reverse: 5′-AGG TTC TGG GCA TTG TGA AC-3′, human PXN (paxillin, NCBI reference sequence: NM_001080855.2), forward: 5′-GGA GTC TAC CAC CTC CCA CA-3′, reverse: 5′-CCA CTG GTC TAA GGG GTC AA-3′, human GAPDH (NCBI reference sequence: NM_001256799.2), forward: 5′-CAT GGG TGT GAA CCA TGA GAA-3′, reverse: 5′-GGT CAT GAG TCC TTC CAC GAT-3′.

Chloramphenicol Acetyltransferase Assay:

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA). .. The primer sequences used were as follows: human PTK2 (FAK, NCBI reference sequence: NM_001199649.1), forward: 5′-TTA TTG GCC ACT GTG GAT GA-3′, reverse: 5′-TAC TCT TGC TGG AGG CTG GT-3′; human TLN1 (Talin1, NCBI reference sequence: NM_006289.3), forward: 5′-TCT CCC AAA ATG CCA AGA AC-3′, reverse: 5′-CTC CAC TAG CCC TTG CTG TC-3′; human VCL (vinculin, NCBI reference sequence: NM_003373.3), forward: 5′-GCC AAG CAG TGC ACA GAT AA-3′, reverse: 5′-AGG TTC TGG GCA TTG TGA AC-3′, human PXN (paxillin, NCBI reference sequence: NM_001080855.2), forward: 5′-GGA GTC TAC CAC CTC CCA CA-3′, reverse: 5′-CCA CTG GTC TAA GGG GTC AA-3′, human GAPDH (NCBI reference sequence: NM_001256799.2), forward: 5′-CAT GGG TGT GAA CCA TGA GAA-3′, reverse: 5′-GGT CAT GAG TCC TTC CAC GAT-3′.

Mouse Assay:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: To examine if the mutation resides in the c-kit gene, total RNA samples were prepared from the skin of adult C57BL/6J and Wadsm/m mice using the Trizol kit following the manufacturer's instructions (Shenergy, Shanghai, China). .. Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI).

Chromatin Immunoprecipitation:

Article Title: Phylogenetic Analysis of Lednice Orthobunyavirus
Article Snippet: Random primed RT-PCR was carried out using a tagged random hexamer [ ] and AMV reverse transcriptase (Promega, Madison, Wisconsin, United States) followed by amplification with DreamTaq DNA polymerase (Thermo Fisher Scientific). .. Sequencing was carried out on Ion Torrent PGM 316 chip using the 200-bp sequencing protocol.

Plasmid Preparation:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI). .. The three fragments were then subcloned into TA cloning vector pMD 18-T (TaKaRa) and sequenced (Bioasia, Shanghai, China).

Electrophoresis:

Article Title: Identification of a Novel Point Mutation of Mouse Proto-Oncogene c-kit Through N-Ethyl-N-nitrosourea Mutagenesis
Article Snippet: PCR products were separated by electrophoresis with 4% agarose II (BioBasic, Markham, ON, Canada) gel. .. Full-length cDNA of c-kit was synthesized with AMV reverse transcriptase (Promega, Madison, WI).

Article Title: Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome
Article Snippet: Single-stranded cDNA was synthesized with AMV reverse transcriptase and random hexamers (Promega). .. PCR products were analyzed by electrophoresis on a 2% agarose gel and bands visualized under UV illumination after ethidium bromide staining.

RNA Extraction:

Article Title: Effect of Dietary Nutrient Density on Small Intestinal Phosphate Transport and Bone Mineralization of Broilers during the Growing Period
Article Snippet: Paragraph title: Total RNA extraction, reverse transcription, and real-time PCR ... Afterward, 1.0 μg of total RNA was reverse-transcribed into single-stranded cDNA with AMV reverse transcriptase and an Oligo (dT)15 primer in the presence of recombinant RNasin ribonuclease inhibitor (A3500; Promega Corporation).

Article Title: LncRNA TP73-AS1 Activates TGF-β1 to Promote the Migration and Invasion of Colorectal Cancer Cell
Article Snippet: Paragraph title: Total RNA Extraction and RT-qPCR ... Following cDNA synthesis using AMV Reverse Transcriptase (Promega Corporation, USA), qPCR reaction systems were prepared using SYBR® Green master mix (Bio-Rad, USA) with 18S rRNA or GAPDH as endogenous controls to detect the expression of TP73-AS1 and TGF-β1, respectively.

Article Title: Reduction of the vitamin D hormonal system in kidney disease is associated with increased renal inflammation
Article Snippet: Total RNA was isolated from frozen kidney biopsies using the GenElute RNA extraction system (Sigma, Poole, UK). .. Aliquots (1.5 µg) of RNA from each biopsy were then reverse transcribed using AMV reverse transcriptase (Promega, Southampton, UK).

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: Paragraph title: Total RNA extraction, reverse transcription, and real-time PCR ... After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.).

Agarose Gel Electrophoresis:

Article Title: Effect of Dietary Nutrient Density on Small Intestinal Phosphate Transport and Bone Mineralization of Broilers during the Growing Period
Article Snippet: The concentration and quality of the extracted RNA were determined through absorbance determination at 260 nm and agarose gel electrophoresis, respectively [ ]. .. Afterward, 1.0 μg of total RNA was reverse-transcribed into single-stranded cDNA with AMV reverse transcriptase and an Oligo (dT)15 primer in the presence of recombinant RNasin ribonuclease inhibitor (A3500; Promega Corporation).

Article Title: Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome
Article Snippet: Single-stranded cDNA was synthesized with AMV reverse transcriptase and random hexamers (Promega). .. PCR products were analyzed by electrophoresis on a 2% agarose gel and bands visualized under UV illumination after ethidium bromide staining.

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: The concentration and quality of the RNA were determined by measuring the absorbance at 260 nm and by agarose gel electrohoresis, respectively ( ). .. After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.).

In Vitro:

Article Title: Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia
Article Snippet: .. In brief, after determining the RNA concentration, in vitro reverse transcription was performed in a volume of 20 μL with a system including 4 μl 25 mM MgCl2 , 2 μL reverse transcription 10×buffer, 2 μL 104 mM dNTP mixture, 0.5 μL ribonuclease inhibitor, 15U AMV reverse transcriptase, 0.5 μg random primers, 1μ total RNA and nuclease-free water to a final volume of 20 μL at 42°C for 15 min and 85°C denaturing (Promega, USA). .. PCR was then performed by using the Ex Taq kit (Takara, Japan) (7.5 μL 2×premix, 10 mM forward and reverse primers, dH2 O to a final volume of 15 μL) in the following conditions: 94°C denature for 3 min, followed by 35 cycles each with 94°C denaturing for 30 s, 58°C annealing for 30 s, and extension for 30 s with PCR Cycler T100 (BioRad, USA).

Incubation:

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Briefly, the cells were seeded to a well of 6-well plate in DMEM containing 100 nM PMA and incubated in a humidified atmosphere of 95% air and 5% CO2 (Forma Scientific, USA) at 37 °C. .. Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA).

Article Title: Dermcidin expression in hepatic cells improves survival without N-glycosylation, but requires asparagine residues
Article Snippet: Reverse transcription was performed in 20 μ l reactions containing 10 μ l of RNA sample plus 10 μ l of RT mix (4 μ l 25 mM MgCl2 , 2 μ l reverse transcription 10 × buffer, 2 μ l dNTP mixture, 1 μ l oligo(dT)15 primer, 0.5 μ l AMV reverse transcriptase and 0.5 μ l recombinant RNasin ribonuclease inhibitor, all from Promega). .. Samples were then incubated in a thermal cycler (PHC-3, Techne) for 1 h at 42°C, followed by 5 min at 99°C.

Article Title: Hypoxia enhances CXCR4 expression in human microvascular endothelial cells and human melanoma cells
Article Snippet: After the appropriate incubation of the endothelial and SK-MEL-5 cells, RNA was isolated with the RNeasy kit (Qiagen, Valencia, CA, USA) with an on-column DNAse treatment (RNAse free DNAse set; Qiagen). .. Next, 1μg of RNA was reverse transcribed using AMV reverse transcriptase (AMV-RT; 0.2 U/μl; Promega, Madison, WI, USA) and its buffer in the presence of oligod(T)16 primers (1 μM; Applied Biosystems, Foster City, CA, USA), 0.2 mM of each of the dNTPs (Sigma-Aldrich) and RNAse inhibitor (RNAsin, 0.4 U/μl; Promega) in a 25 μl reaction mixture.

Concentration Assay:

Article Title: Effect of Dietary Nutrient Density on Small Intestinal Phosphate Transport and Bone Mineralization of Broilers during the Growing Period
Article Snippet: The concentration and quality of the extracted RNA were determined through absorbance determination at 260 nm and agarose gel electrophoresis, respectively [ ]. .. Afterward, 1.0 μg of total RNA was reverse-transcribed into single-stranded cDNA with AMV reverse transcriptase and an Oligo (dT)15 primer in the presence of recombinant RNasin ribonuclease inhibitor (A3500; Promega Corporation).

Article Title: The Influence of Dietary Calcium and Phosphorus Imbalance on Intestinal NaPi-IIb and Calbindin mRNA Expression and Tibia Parameters of Broilers
Article Snippet: The concentration and quality of the RNA were determined by measuring the absorbance at 260 nm and by agarose gel electrohoresis, respectively ( ). .. After extraction, 1.0 μg of total RNA was made into single-stranded cDNA with AMV Reverse Transcriptase using an Oligo (dT)15 primer in the presence of Recombinant RNasin Ribonuclease Inhibitor (A3500, Promega Corp.).

Article Title: Role of LAMA4 Gene in Regulating Extravillous Trophoblasts in Pathogenesis of Preeclampsia
Article Snippet: .. In brief, after determining the RNA concentration, in vitro reverse transcription was performed in a volume of 20 μL with a system including 4 μl 25 mM MgCl2 , 2 μL reverse transcription 10×buffer, 2 μL 104 mM dNTP mixture, 0.5 μL ribonuclease inhibitor, 15U AMV reverse transcriptase, 0.5 μg random primers, 1μ total RNA and nuclease-free water to a final volume of 20 μL at 42°C for 15 min and 85°C denaturing (Promega, USA). .. PCR was then performed by using the Ex Taq kit (Takara, Japan) (7.5 μL 2×premix, 10 mM forward and reverse primers, dH2 O to a final volume of 15 μL) in the following conditions: 94°C denature for 3 min, followed by 35 cycles each with 94°C denaturing for 30 s, 58°C annealing for 30 s, and extension for 30 s with PCR Cycler T100 (BioRad, USA).

CTG Assay:

Article Title: Priming stem cells with protein kinase C activator enhances early stem cell-chondrocyte interaction by increasing adhesion molecules
Article Snippet: Complementary DNA (cDNA) was synthesized from RNA by AMV reverse transcriptase in provided in a RT system kit (Promega, Fitchburg, WI, USA). .. The primer sequences used were as follows: human PTK2 (FAK, NCBI reference sequence: NM_001199649.1), forward: 5′-TTA TTG GCC ACT GTG GAT GA-3′, reverse: 5′-TAC TCT TGC TGG AGG CTG GT-3′; human TLN1 (Talin1, NCBI reference sequence: NM_006289.3), forward: 5′-TCT CCC AAA ATG CCA AGA AC-3′, reverse: 5′-CTC CAC TAG CCC TTG CTG TC-3′; human VCL (vinculin, NCBI reference sequence: NM_003373.3), forward: 5′-GCC AAG CAG TGC ACA GAT AA-3′, reverse: 5′-AGG TTC TGG GCA TTG TGA AC-3′, human PXN (paxillin, NCBI reference sequence: NM_001080855.2), forward: 5′-GGA GTC TAC CAC CTC CCA CA-3′, reverse: 5′-CCA CTG GTC TAA GGG GTC AA-3′, human GAPDH (NCBI reference sequence: NM_001256799.2), forward: 5′-CAT GGG TGT GAA CCA TGA GAA-3′, reverse: 5′-GGT CAT GAG TCC TTC CAC GAT-3′.

Staining:

Article Title: Dysfunctional SEMA3E signaling underlies gonadotropin-releasing hormone neuron deficiency in Kallmann syndrome
Article Snippet: Single-stranded cDNA was synthesized with AMV reverse transcriptase and random hexamers (Promega). .. PCR products were analyzed by electrophoresis on a 2% agarose gel and bands visualized under UV illumination after ethidium bromide staining.

Nanopore Sequencing:

Article Title: Molecular Identification of a Novel Hantavirus in Malaysian Bronze Tube-Nosed Bats (Murina aenea)
Article Snippet: Paragraph title: 2.3. Metagenomic cDNA Preparation and Nanopore Sequencing ... Samples were then subjected to the Sequence Independent Single Primer Amplification (SISPA) approach [ ]. cDNA amplification was performed by an AMV Reverse Transcriptase (Promega) according to the provided manual by the manufacturers using the FR26RV-N (5′-GCCGGAGCTCTGCAGATATCNNNNNN-3′) primer.

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  • 99
    Promega avian myeloblastosis virus reverse transcriptase
    Primer extension analysis of human brain RNA. Oligonucleotides GSP-6, PEXT1 and PEXT2 flanking the 5′ termini of the human LGA cDNA clone were 32 P end-labeled with [γ- 32 P]ATP and T4 polynucleotide kinase. Poly(A) + mRNA of human brain (approx. 0.5–1 µg) was mixed with the radioactive oligonucleotides and reverse-transcribed with avian <t>myeloblastosis</t> virus reverse transcriptase; the products from primer extension reactions were separated by electrophoresis on an 8% (w/v) denaturing polyacrylamide gel. Lanes are labeled with the name of the specific primer used in each case. The size of the single-stranded cDNA products (in bases) was estimated approximately by comparison with ФX174 HinfI DNA markers shown at the left in lane M. C+ lane: product amplified with a positive control of kanamycin. C- lanes: negative controls using PEXT2 with mRNA omitted. The black arrow indicates the specific GA products of approx. 140 bases obtained by reverse transcription with primer PEXT2. Top panel: scheme showing the approximate position of the three primers with regard to the TSS (+1) and the two first ATG codons of human LGA transcript.
    Avian Myeloblastosis Virus Reverse Transcriptase, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/avian myeloblastosis virus reverse transcriptase/product/Promega
    Average 99 stars, based on 98 article reviews
    Price from $9.99 to $1999.99
    avian myeloblastosis virus reverse transcriptase - by Bioz Stars, 2020-02
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    90
    Promega amv reverse transcriptase
    Encapsidation of <t>pgRNA.</t> Primer extension using oligo 1948− extended with avian myeloblastosis virus <t>(AMV)</t> reverse transcriptase (RT) was used to measure the level of 5′ ends of encapsidated and reference RNAs. (A and B) Representative gels used to measure the encapsidation efficiency of total and encapsidated pgRNA, respectively. In each gel, the positions of the pgRNA and reference RNA (ref. RNA) are indicated. A sequencing ladder is provided for reference. (C) Histogram of the efficiency of encapsidation of the 5′ end of pgRNA, with standard deviations of normalized values represented by error bars. (D) Formula used to calculate encapsidation. An asterisk (*) indicates a significant difference from the WT reference ( P
    Amv Reverse Transcriptase, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 248 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/amv reverse transcriptase/product/Promega
    Average 90 stars, based on 248 article reviews
    Price from $9.99 to $1999.99
    amv reverse transcriptase - by Bioz Stars, 2020-02
    90/100 stars
      Buy from Supplier

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    Primer extension analysis of human brain RNA. Oligonucleotides GSP-6, PEXT1 and PEXT2 flanking the 5′ termini of the human LGA cDNA clone were 32 P end-labeled with [γ- 32 P]ATP and T4 polynucleotide kinase. Poly(A) + mRNA of human brain (approx. 0.5–1 µg) was mixed with the radioactive oligonucleotides and reverse-transcribed with avian myeloblastosis virus reverse transcriptase; the products from primer extension reactions were separated by electrophoresis on an 8% (w/v) denaturing polyacrylamide gel. Lanes are labeled with the name of the specific primer used in each case. The size of the single-stranded cDNA products (in bases) was estimated approximately by comparison with ФX174 HinfI DNA markers shown at the left in lane M. C+ lane: product amplified with a positive control of kanamycin. C- lanes: negative controls using PEXT2 with mRNA omitted. The black arrow indicates the specific GA products of approx. 140 bases obtained by reverse transcription with primer PEXT2. Top panel: scheme showing the approximate position of the three primers with regard to the TSS (+1) and the two first ATG codons of human LGA transcript.

    Journal: PLoS ONE

    Article Title: Mammalian Glutaminase Gls2 Gene Encodes Two Functional Alternative Transcripts by a Surrogate Promoter Usage Mechanism

    doi: 10.1371/journal.pone.0038380

    Figure Lengend Snippet: Primer extension analysis of human brain RNA. Oligonucleotides GSP-6, PEXT1 and PEXT2 flanking the 5′ termini of the human LGA cDNA clone were 32 P end-labeled with [γ- 32 P]ATP and T4 polynucleotide kinase. Poly(A) + mRNA of human brain (approx. 0.5–1 µg) was mixed with the radioactive oligonucleotides and reverse-transcribed with avian myeloblastosis virus reverse transcriptase; the products from primer extension reactions were separated by electrophoresis on an 8% (w/v) denaturing polyacrylamide gel. Lanes are labeled with the name of the specific primer used in each case. The size of the single-stranded cDNA products (in bases) was estimated approximately by comparison with ФX174 HinfI DNA markers shown at the left in lane M. C+ lane: product amplified with a positive control of kanamycin. C- lanes: negative controls using PEXT2 with mRNA omitted. The black arrow indicates the specific GA products of approx. 140 bases obtained by reverse transcription with primer PEXT2. Top panel: scheme showing the approximate position of the three primers with regard to the TSS (+1) and the two first ATG codons of human LGA transcript.

    Article Snippet: 0.5–1 µg) was mixed with the radioactive oligonucleotides and reverse transcribed with avian myeloblastosis virus reverse transcriptase, with the use of the protocol provided by the manufacturer (Primer Extension Analysis kit; Promega).

    Techniques: Labeling, Electrophoresis, Amplification, Positive Control

    FcRn expression in normal adult human small intestine and human intestinal epithelial cell lines. Western blots of total cellular protein (13 μg protein per lane, a ; 10 μg protein per lane, b ) isolated from the indicated source using affinity-purified rabbit antisera raised against amino acids 112–125 ( a ) or amino acids 174–188 ( b ). ( c ) RT-PCR detection of FcγRI transcripts. Total RNA (2 μg) from T84 (lanes 3 and 4), MOLT-4 (lanes 5 and 6; negative control), and U937 (lanes 1 and 2; positive control) cell lines was incubated with an oligo-dT primer with (odd-numbered lanes) or without (even-numbered lanes) avian myeloblastosis virus–RT (AMV-RT), and a nested PCR was performed with primers specific for FcγRI cDNA (top) or for β-actin (bottom).

    Journal: Journal of Clinical Investigation

    Article Title: Bidirectional FcRn-dependent IgG transport in a polarized human intestinal epithelial cell line

    doi:

    Figure Lengend Snippet: FcRn expression in normal adult human small intestine and human intestinal epithelial cell lines. Western blots of total cellular protein (13 μg protein per lane, a ; 10 μg protein per lane, b ) isolated from the indicated source using affinity-purified rabbit antisera raised against amino acids 112–125 ( a ) or amino acids 174–188 ( b ). ( c ) RT-PCR detection of FcγRI transcripts. Total RNA (2 μg) from T84 (lanes 3 and 4), MOLT-4 (lanes 5 and 6; negative control), and U937 (lanes 1 and 2; positive control) cell lines was incubated with an oligo-dT primer with (odd-numbered lanes) or without (even-numbered lanes) avian myeloblastosis virus–RT (AMV-RT), and a nested PCR was performed with primers specific for FcγRI cDNA (top) or for β-actin (bottom).

    Article Snippet: RNA (2 μg) was reverse-transcribed to cDNA with an oligo-dT primer (Promega Corp., Madison, Wisconsin, USA) and avian myeloblastosis virus reverse transcriptase (Promega Corp.).

    Techniques: Expressing, Western Blot, Isolation, Affinity Purification, Reverse Transcription Polymerase Chain Reaction, Negative Control, Positive Control, Incubation, Nested PCR

    Encapsidation of pgRNA. Primer extension using oligo 1948− extended with avian myeloblastosis virus (AMV) reverse transcriptase (RT) was used to measure the level of 5′ ends of encapsidated and reference RNAs. (A and B) Representative gels used to measure the encapsidation efficiency of total and encapsidated pgRNA, respectively. In each gel, the positions of the pgRNA and reference RNA (ref. RNA) are indicated. A sequencing ladder is provided for reference. (C) Histogram of the efficiency of encapsidation of the 5′ end of pgRNA, with standard deviations of normalized values represented by error bars. (D) Formula used to calculate encapsidation. An asterisk (*) indicates a significant difference from the WT reference ( P

    Journal: Journal of Virology

    Article Title: The Arginine Clusters of the Carboxy-Terminal Domain of the Core Protein of Hepatitis B Virus Make Pleiotropic Contributions to Genome Replication ▿

    doi: 10.1128/JVI.01957-10

    Figure Lengend Snippet: Encapsidation of pgRNA. Primer extension using oligo 1948− extended with avian myeloblastosis virus (AMV) reverse transcriptase (RT) was used to measure the level of 5′ ends of encapsidated and reference RNAs. (A and B) Representative gels used to measure the encapsidation efficiency of total and encapsidated pgRNA, respectively. In each gel, the positions of the pgRNA and reference RNA (ref. RNA) are indicated. A sequencing ladder is provided for reference. (C) Histogram of the efficiency of encapsidation of the 5′ end of pgRNA, with standard deviations of normalized values represented by error bars. (D) Formula used to calculate encapsidation. An asterisk (*) indicates a significant difference from the WT reference ( P

    Article Snippet: For simultaneous detection of RNA and DNA by primer extension (as shown in Fig. and ) a cDNA of pgRNA was generated using AMV reverse transcriptase (Promega) and an unlabeled primer.

    Techniques: Sequencing

    Utilization of dGDP as substrate by the AMV RT and T4 DNA polymerase Sequences of the DNA/RNA or DNA/DNA hybrid substrates are shown above the gels. The templates of the substrates specify incorporation for a dA and a dG residue. The enzymes were assayed with 0.165 μM 32 P‐dATP (asterisk) and either dGMP, dGDP, or dGTP at 100 μM.

    Journal: The EMBO Journal

    Article Title: A single nucleotide incorporation step limits human telomerase repeat addition activity

    doi: 10.15252/embj.201797953

    Figure Lengend Snippet: Utilization of dGDP as substrate by the AMV RT and T4 DNA polymerase Sequences of the DNA/RNA or DNA/DNA hybrid substrates are shown above the gels. The templates of the substrates specify incorporation for a dA and a dG residue. The enzymes were assayed with 0.165 μM 32 P‐dATP (asterisk) and either dGMP, dGDP, or dGTP at 100 μM.

    Article Snippet: One microliter of RRL reconstituted telomerase enzyme, 1 unit of AMV RT (Promega), 0.5 units of Taq DNA pol III (NEB), 1 unit of T4 DNA pol (Fermentas), or 0.5 units of Klenow fragment of DNA pol I (Invitrogen) were assayed in 10 μl reactions containing 1× telomerase reaction buffer, 40 μM of denoted pre‐annealed DNA/RNA or DNA/DNA hybrid substrates, 100 μM dGTP, dGDP, or dGMP, and 0.165 μM α‐32 P‐dATP.

    Techniques: