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Promega dntp
Dntp, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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dntp - by Bioz Stars, 2020-02
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Related Articles

Centrifugation:

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: After incubation at –20° C overnight, RNA was precipitated by centrifugation at14 K rpm for 30 min at 4o C and the precipitated RNA was washed twice with 70% ethanol. .. The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor.

Amplification:

Article Title: β-Thalassaemia Major in a Spanish Patient due to a Compound Heterozygosity for CD39 C - > T/-28 A - > C
Article Snippet: The full coding, the 5′UTR, and the 3′UTR sequences of β -globin gene (GenBank accession no. U01317) were amplified and sequenced to discard other mutations. .. The amplifications were performed in a 25 μ L volume, containing 25 mM Cl2 Mg, 200 μ M of each dNTP (Promega, Madrid, Spain), 2.5 U Expand Long Template PCR System (Roche), 0.6 μ M each primer, and 100 ng DNA.

Article Title: The biocontrol bacterium Pseudomonas fluorescens Pf29Arp strain affects the pathogenesis-related gene expression of the take-all fungus Gaeumannomyces graminis var. tritici on wheat roots
Article Snippet: These selected amplified sequences, Wn1T7 and Wn2T7 , were verified by DNA sequencing (GATC Biotech., Mulhouse, France) and used as templates for in vitro transcription with the MEGAscript Kit (Ambion, Austin, TX, USA) to obtain purified Wn1 and Wn2 RNA. .. RT was carried out in 20-µL volumes with 500 ng of RNA, 1 µg of oligo-dT primers (random primers for bacterial RNA), 1 × ImPromII reaction buffer, 3 mm MgCl2 , 0.5 mm of each dNTP, 20 U of RNasin Ribonuclease Inhibitor and 1 µL of ImProm-II™ (Promega Corp.).

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: The primers used for PCR amplification of 16S ribosomal DNA sequences were the universal primers 16SP0 ( 5′-GAAGAGTTTGATCCTGGCTCAG-3′ ) and 16SP6 ( 5′-CTACGGCTACCTTGTTACGA-3′ ). .. The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega).

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: .. PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France). .. Sense F1 (5'- TCTGACATGAGAGAGGCCAACTAC-3') and anti-sense R1 (5'-GTCAGGCAGGCCACGAGGTCT-3') primers were designed on the basis of our porcine SAA cDNA sequence obtained from the F0-R0 PCR product.

Article Title: A second generation framework for the analysis of microsatellites in expressed sequence tags and the development of EST-SSR markers for a conifer, Cryptomeria japonica
Article Snippet: 5 ng genomic DNA, 1 × PCR buffer, 200 μM of each dNTP, 1.5 mM MgCl2 , 0.2 μM of each synthesized primer, and 0.25 U of Taq polymerase (Promega Madison, USA), using the following program: 94°C for 5 min, then 40 cycles of 94°C for 30 s, 55–62°C for 30 s and 72°C for 30 s, followed by a final extension at 72°C for 5 min. .. The PCR products were electrophoretically separated on 2% agarose gels and stained with ethidium bromide to check for successful amplification.

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: .. The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor. ..

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: The second region was a ~658 nucleotide portion of the cytochrome c oxidase subunit I (COI ) gene amplified using slightly degenerate versions of standard primers [ ]. .. Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Article Title: Multiple Signaling Pathways Are Involved in the Interleukine-4 Regulated Expression of DC-SIGN in THP-1 Cell Line
Article Snippet: And the DC-SIGN promoter without Ets-1 binding site was amplified in the same way, using primers of P1, P2, P6, P7, and P8, as shown in . .. The mixture of PCR reaction consisted of 0.2 μ L DNA template, 2 μ L forward/reverse primers (10 mM), 4 μ L MgCl2 (25 mM), 4 μ L dNTP (2.5 mM), 5 μ L 10×PCR buffer, 0.5 μ L Pfu DNA polymerase (2 U/μ L, promega), and the final volume was taken to 50 μ L with water.

Synthesized:

Article Title: Taeyeumjoweetang Affects Body Weight and Obesity-related Genes in Mice
Article Snippet: .. The reaction used a reaction solution of total RNA (3–5 μg), oligo d (T) 12–18 (1 μg), 2 μl dNTP (10 mM), MMLV reverse transcriptase (200 U), DTT (10 mM), RNase inhibitor (1 μl; Promega, USA) and a 20 μl buffer solution (50 mM Tris–Cl, pH 8.3, 75 mM KCl, 3 mM MgCl2 ) at 42°C for 60 min, after which cDNA was synthesized. .. Real-time reverse transcription polymerase chain reaction was done by mixing cDNA (diluted 10 times) with 2× SYBR-Green buffer solution (Roche Diagnostics Ltd, UK), which contained reverse transcription enzymes, using a LightCycler rapid thermal cycler system (Roche).

Article Title: A second generation framework for the analysis of microsatellites in expressed sequence tags and the development of EST-SSR markers for a conifer, Cryptomeria japonica
Article Snippet: .. 5 ng genomic DNA, 1 × PCR buffer, 200 μM of each dNTP, 1.5 mM MgCl2 , 0.2 μM of each synthesized primer, and 0.25 U of Taq polymerase (Promega Madison, USA), using the following program: 94°C for 5 min, then 40 cycles of 94°C for 30 s, 55–62°C for 30 s and 72°C for 30 s, followed by a final extension at 72°C for 5 min. .. The PCR products were electrophoretically separated on 2% agarose gels and stained with ethidium bromide to check for successful amplification.

Construct:

Article Title: Selective stalling of human translation through small-molecule engagement of the ribosome nascent chain
Article Snippet: Translation reactions were transferred to ice and supplemented with 0.5 mM each dNTP, 50 mM Tris-Cl pH 8.3, 9.4 mM MgCl2 , 10 mM DTT, 0.8 U/μL RNasin Plus (Promega), and 0.1 mg/mL cycloheximide. .. The primer used for full-length PCSK9 constructs was 5′-ATCTTGGTGAGGTATCCCCG-3′.

SYBR Green Assay:

Article Title: Taeyeumjoweetang Affects Body Weight and Obesity-related Genes in Mice
Article Snippet: The reaction used a reaction solution of total RNA (3–5 μg), oligo d (T) 12–18 (1 μg), 2 μl dNTP (10 mM), MMLV reverse transcriptase (200 U), DTT (10 mM), RNase inhibitor (1 μl; Promega, USA) and a 20 μl buffer solution (50 mM Tris–Cl, pH 8.3, 75 mM KCl, 3 mM MgCl2 ) at 42°C for 60 min, after which cDNA was synthesized. .. Real-time reverse transcription polymerase chain reaction was done by mixing cDNA (diluted 10 times) with 2× SYBR-Green buffer solution (Roche Diagnostics Ltd, UK), which contained reverse transcription enzymes, using a LightCycler rapid thermal cycler system (Roche).

Incubation:

Article Title: GmPHD5 acts as an important regulator for crosstalk between histone H3K4 di-methylation and H3K14 acetylation in response to salinity stress in soybean
Article Snippet: Immuno-precipitating antibody was then added and the mixture was incubated overnight with rotation at 4°C. .. ChIP-PCR reactions were set up as follows: 4 ul template (~ < 0.1 nmol) was mixed with 0.4 ul dNTP (10 mM), 0.4 ul forward primer (10 uM), 0.4 ul reverse primer (10 uM), 2 ul 10 × PCR buffer, 0.25 ul Taq polymerase (Promega, Wisconsin, USA), and 1 ul MgCl2 (25 mM).

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: After incubation at –20° C overnight, RNA was precipitated by centrifugation at14 K rpm for 30 min at 4o C and the precipitated RNA was washed twice with 70% ethanol. .. The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor.

Article Title: A quantitative assay for measuring mRNA decapping by splinted ligation reverse transcription polymerase chain reaction: qSL-RT-PCR
Article Snippet: .. Reverse transcription reactions contained 20 picomoles of gene-specific reverse primer (NB39 for RPL41A or NB56 for YLR084C), 0.5 mM each dNTP, 1.2 mM MgCl2 , 1 x GoScript buffer, and 1 μL of GoScript (Promega) in a 20-μL reaction and incubated 1 h at 42°C, then heat inactivated for 15 min at 65°C. .. Amplification of PCR products was measured using GoTaq qPCR master mix (Promega) with 200 nM of each primer in 50-μL reactions.

Luciferase:

Article Title: Multiple Signaling Pathways Are Involved in the Interleukine-4 Regulated Expression of DC-SIGN in THP-1 Cell Line
Article Snippet: Paragraph title: 2.6. Construction of DC-SIGN Promoter Luciferase Reporter Plasmids and the Activity Detection ... The mixture of PCR reaction consisted of 0.2 μ L DNA template, 2 μ L forward/reverse primers (10 mM), 4 μ L MgCl2 (25 mM), 4 μ L dNTP (2.5 mM), 5 μ L 10×PCR buffer, 0.5 μ L Pfu DNA polymerase (2 U/μ L, promega), and the final volume was taken to 50 μ L with water.

Activity Assay:

Article Title: Multiple Signaling Pathways Are Involved in the Interleukine-4 Regulated Expression of DC-SIGN in THP-1 Cell Line
Article Snippet: Paragraph title: 2.6. Construction of DC-SIGN Promoter Luciferase Reporter Plasmids and the Activity Detection ... The mixture of PCR reaction consisted of 0.2 μ L DNA template, 2 μ L forward/reverse primers (10 mM), 4 μ L MgCl2 (25 mM), 4 μ L dNTP (2.5 mM), 5 μ L 10×PCR buffer, 0.5 μ L Pfu DNA polymerase (2 U/μ L, promega), and the final volume was taken to 50 μ L with water.

Expressing:

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega). .. RT-qPCR was performed using proprietary TaqMan® Gene Expression Assay primers exclusive to human ASCT2, LAT1, alanine-serine-cysteine transporter 1 (ASCT1), sodium-dependent neutral amino acid transporter 2 (SNAT2), large neutral amino acid transporter 2 (LAT2), glutamine synthetase (GLUL), and cystine-glutamate transporter (xCT) mRNA transcripts, and all measurements were normalized to two housekeeping genes (HKG): TATA-binding protein (TBP) and hydroxymethylbilane synthase (HMBS) (all TaqMan probes were from Applied Biosystems, Foster City, CA, USA).

Modification:

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: This region was amplified in two overlapping portions using the primers 12Sai [5'-AAACTAGGATTAGATACCCTATTAT-3'; 63] with 16Srgast [5'-GCCATGATGCAAAAGGTAC-3'], and 12Sf [5'-GCACACATCGCCCGTCGCTCT-3'; reverse complement of 12Sb' in 63] with 16SbrH-alt [5'-CCGGTCTGAACTCAGATCATGT-3'; slight modification of 16Sbr in 63]. .. Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Real-time Polymerase Chain Reaction:

Article Title: β-Thalassaemia Major in a Spanish Patient due to a Compound Heterozygosity for CD39 C - > T/-28 A - > C
Article Snippet: 2.3. β -Globin Gene Analysis The propositus and his parents were studied for the most frequent β -thalassaemia mutations in the Mediterranean area by procedures based upon real-time PCR and specific fluorescent labeled hybridization probes including: IVSI-II-745 C → G, CD5 (-CT), CD6 (-A), CD8 (-AA), CD39 C → T, CD37 G → A, IVSI-I G → A, IVSI-6 T → C, and IVSI-110 G → A [ , ]. .. The amplifications were performed in a 25 μ L volume, containing 25 mM Cl2 Mg, 200 μ M of each dNTP (Promega, Madrid, Spain), 2.5 U Expand Long Template PCR System (Roche), 0.6 μ M each primer, and 100 ng DNA.

Article Title: The biocontrol bacterium Pseudomonas fluorescens Pf29Arp strain affects the pathogenesis-related gene expression of the take-all fungus Gaeumannomyces graminis var. tritici on wheat roots
Article Snippet: RT was carried out in 20-µL volumes with 500 ng of RNA, 1 µg of oligo-dT primers (random primers for bacterial RNA), 1 × ImPromII reaction buffer, 3 mm MgCl2 , 0.5 mm of each dNTP, 20 U of RNasin Ribonuclease Inhibitor and 1 µL of ImProm-II™ (Promega Corp.). .. Melting curve analysis was performed after the qPCR to confirm that the signal, obtained only in the Wn1 and Wn2 RNA samples, was the result of a single product of amplification and not caused by primer dimers or an arbitrary amplification.

Hybridization:

Article Title: β-Thalassaemia Major in a Spanish Patient due to a Compound Heterozygosity for CD39 C - > T/-28 A - > C
Article Snippet: 2.3. β -Globin Gene Analysis The propositus and his parents were studied for the most frequent β -thalassaemia mutations in the Mediterranean area by procedures based upon real-time PCR and specific fluorescent labeled hybridization probes including: IVSI-II-745 C → G, CD5 (-CT), CD6 (-A), CD8 (-AA), CD39 C → T, CD37 G → A, IVSI-I G → A, IVSI-6 T → C, and IVSI-110 G → A [ , ]. .. The amplifications were performed in a 25 μ L volume, containing 25 mM Cl2 Mg, 200 μ M of each dNTP (Promega, Madrid, Spain), 2.5 U Expand Long Template PCR System (Roche), 0.6 μ M each primer, and 100 ng DNA.

Ligation:

Article Title: A quantitative assay for measuring mRNA decapping by splinted ligation reverse transcription polymerase chain reaction: qSL-RT-PCR
Article Snippet: For splinted ligation samples, this step is incorporated into the Splinted Ligation method described above. .. Reverse transcription reactions contained 20 picomoles of gene-specific reverse primer (NB39 for RPL41A or NB56 for YLR084C), 0.5 mM each dNTP, 1.2 mM MgCl2 , 1 x GoScript buffer, and 1 μL of GoScript (Promega) in a 20-μL reaction and incubated 1 h at 42°C, then heat inactivated for 15 min at 65°C.

Cell Culture:

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: RNA Isolation and RT-qPCR All cell lines were cultured in 150 mm culture dish; when the cells reached 80–90% confluence, an RNA isolation was performed using TRIzol® reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer instructions, and this was followed by an alcohol precipitation to concentrate the samples. .. One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega).

Generated:

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega). .. Although no new sequence data was generated, all new data has been deposited in GenBank.

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega). .. The 2−ΔΔC t method was used to calculate the fold difference in expression relative to nonsilencing (NS) controls generated from each parent cell line.

Polymerase Chain Reaction:

Article Title: GmPHD5 acts as an important regulator for crosstalk between histone H3K4 di-methylation and H3K14 acetylation in response to salinity stress in soybean
Article Snippet: .. ChIP-PCR reactions were set up as follows: 4 ul template (~ < 0.1 nmol) was mixed with 0.4 ul dNTP (10 mM), 0.4 ul forward primer (10 uM), 0.4 ul reverse primer (10 uM), 2 ul 10 × PCR buffer, 0.25 ul Taq polymerase (Promega, Wisconsin, USA), and 1 ul MgCl2 (25 mM). ..

Article Title: β-Thalassaemia Major in a Spanish Patient due to a Compound Heterozygosity for CD39 C - > T/-28 A - > C
Article Snippet: .. The amplifications were performed in a 25 μ L volume, containing 25 mM Cl2 Mg, 200 μ M of each dNTP (Promega, Madrid, Spain), 2.5 U Expand Long Template PCR System (Roche), 0.6 μ M each primer, and 100 ng DNA. .. Amplification was performed in a PTC-100 thermal cycler (MJ Research INC, Madrid, Spain) for one cycle of 10 minutes at 95°C and then for 35 cycles of 20 seconds at 95°C, 20 seconds at 62°C, and 1 minute at 72°C, followed by one final cycle similar to the previous one but with 10 minutes at 72°C.

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: .. The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega). .. The PCR product was purified using GFX PCR DNA and a Gel Band Kit (GE Healthcare), then sequenced using Big Dye v3.1 Kit and 3730 DNA Analyser (Applied Biosystems).

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: .. PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France). .. Sense F1 (5'- TCTGACATGAGAGAGGCCAACTAC-3') and anti-sense R1 (5'-GTCAGGCAGGCCACGAGGTCT-3') primers were designed on the basis of our porcine SAA cDNA sequence obtained from the F0-R0 PCR product.

Article Title: No association of the insulin gene VNTR polymorphism with polycystic ovary syndrome in a Han Chinese population
Article Snippet: .. The PCR reaction mixture contained 50 ng genomic DNA, 0.5 μM of each primer (forward: 5'-AGC AGG TCT GTT CCA AGG-3' and reverse: 5'-CTT GGG TGT GTA GAA GAA GC-3'), 10 mM Tris-HCl (pH 9.0), 50 mM KCl, 1.5 mM MgCl2, 0.1% Triton X-100, 0.2 mM of each dNTP, and 0.75 unit Tag DNA polymerase (Promega, USA) in a final volume of 25 ml. ..

Article Title: Genomic Fossils Calibrate the Long-Term Evolution of Hepadnaviruses
Article Snippet: .. PCR mix was buffer (5×), 5 µl; MgCl2 (25 mM), 2 µl; dNTP (10 mM), 0.5 µl; primer 1 (10 µM), 1 µl; primer 2 (10 µM), 1 µl; Taq (GoTaq, Promega), 1.25 U; DNA, 30–100 ng; and H2 O up to 25 µl. .. PCR products were directly sequenced on an ABI 3130XL sequencer (Applied Biosystems).

Article Title: A second generation framework for the analysis of microsatellites in expressed sequence tags and the development of EST-SSR markers for a conifer, Cryptomeria japonica
Article Snippet: .. 5 ng genomic DNA, 1 × PCR buffer, 200 μM of each dNTP, 1.5 mM MgCl2 , 0.2 μM of each synthesized primer, and 0.25 U of Taq polymerase (Promega Madison, USA), using the following program: 94°C for 5 min, then 40 cycles of 94°C for 30 s, 55–62°C for 30 s and 72°C for 30 s, followed by a final extension at 72°C for 5 min. .. The PCR products were electrophoretically separated on 2% agarose gels and stained with ethidium bromide to check for successful amplification.

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: The complimentary DNA (cDNA) and the first PCR reaction of the nested PCR detection system for the HCV RNA was performed in a 50 µL volume single-step reaction using the Ready-To-Go RT-PCR beads (Healthcare Life Sciences, USA), 10 µM from each of the RT downstream primer, PCR forward primer and reverse primer P2. .. The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor.

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: The polymerase chain reaction (PCR) was used to amplify two portions of the mitochondrial genome. .. Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Article Title: Multiple Signaling Pathways Are Involved in the Interleukine-4 Regulated Expression of DC-SIGN in THP-1 Cell Line
Article Snippet: .. The mixture of PCR reaction consisted of 0.2 μ L DNA template, 2 μ L forward/reverse primers (10 mM), 4 μ L MgCl2 (25 mM), 4 μ L dNTP (2.5 mM), 5 μ L 10×PCR buffer, 0.5 μ L Pfu DNA polymerase (2 U/μ L, promega), and the final volume was taken to 50 μ L with water. .. PCR was performed for 30 cycles of denaturation (95°C, 30 s), annealing (57–62°C, 1 min), and extension (72°C, 1 min).

DNA Sequencing:

Article Title: The biocontrol bacterium Pseudomonas fluorescens Pf29Arp strain affects the pathogenesis-related gene expression of the take-all fungus Gaeumannomyces graminis var. tritici on wheat roots
Article Snippet: These selected amplified sequences, Wn1T7 and Wn2T7 , were verified by DNA sequencing (GATC Biotech., Mulhouse, France) and used as templates for in vitro transcription with the MEGAscript Kit (Ambion, Austin, TX, USA) to obtain purified Wn1 and Wn2 RNA. .. RT was carried out in 20-µL volumes with 500 ng of RNA, 1 µg of oligo-dT primers (random primers for bacterial RNA), 1 × ImPromII reaction buffer, 3 mm MgCl2 , 0.5 mm of each dNTP, 20 U of RNasin Ribonuclease Inhibitor and 1 µL of ImProm-II™ (Promega Corp.).

Sequencing:

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega). .. Although no new sequence data was generated, all new data has been deposited in GenBank.

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: As no homologous porcine EST was identified based on the porcine SAA cDNA sequence, primers F0 et R0 were designed from dog, mink, guinea pig, rabbit and human SAA cDNA sequences, CM59173, in a region relatively well conserved between SAA of different subtypes and different animal origins, using VectorNTI software (Invitrogen, Cergy Pontoise, France). .. PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France).

Article Title: Genomic Fossils Calibrate the Long-Term Evolution of Hepadnaviruses
Article Snippet: Paragraph title: PCR and Sequencing ... PCR mix was buffer (5×), 5 µl; MgCl2 (25 mM), 2 µl; dNTP (10 mM), 0.5 µl; primer 1 (10 µM), 1 µl; primer 2 (10 µM), 1 µl; Taq (GoTaq, Promega), 1.25 U; DNA, 30–100 ng; and H2 O up to 25 µl.

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: Paragraph title: Sampling, DNA extraction, and sequencing ... Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Binding Assay:

Article Title: Multiple Signaling Pathways Are Involved in the Interleukine-4 Regulated Expression of DC-SIGN in THP-1 Cell Line
Article Snippet: And the DC-SIGN promoter without Ets-1 binding site was amplified in the same way, using primers of P1, P2, P6, P7, and P8, as shown in . .. The mixture of PCR reaction consisted of 0.2 μ L DNA template, 2 μ L forward/reverse primers (10 mM), 4 μ L MgCl2 (25 mM), 4 μ L dNTP (2.5 mM), 5 μ L 10×PCR buffer, 0.5 μ L Pfu DNA polymerase (2 U/μ L, promega), and the final volume was taken to 50 μ L with water.

DNA Extraction:

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: Paragraph title: Sampling, DNA extraction, and sequencing ... Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Isolation:

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: DNA was isolated from ~50 mg of tissue using standard phenol/chloroform methods [ ], or Qiagen's Dneasy extraction kit. .. Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: Paragraph title: 4.4. RNA Isolation and RT-qPCR ... One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega).

Labeling:

Article Title: β-Thalassaemia Major in a Spanish Patient due to a Compound Heterozygosity for CD39 C - > T/-28 A - > C
Article Snippet: 2.3. β -Globin Gene Analysis The propositus and his parents were studied for the most frequent β -thalassaemia mutations in the Mediterranean area by procedures based upon real-time PCR and specific fluorescent labeled hybridization probes including: IVSI-II-745 C → G, CD5 (-CT), CD6 (-A), CD8 (-AA), CD39 C → T, CD37 G → A, IVSI-I G → A, IVSI-6 T → C, and IVSI-110 G → A [ , ]. .. The amplifications were performed in a 25 μ L volume, containing 25 mM Cl2 Mg, 200 μ M of each dNTP (Promega, Madrid, Spain), 2.5 U Expand Long Template PCR System (Roche), 0.6 μ M each primer, and 100 ng DNA.

Purification:

Article Title: The biocontrol bacterium Pseudomonas fluorescens Pf29Arp strain affects the pathogenesis-related gene expression of the take-all fungus Gaeumannomyces graminis var. tritici on wheat roots
Article Snippet: These selected amplified sequences, Wn1T7 and Wn2T7 , were verified by DNA sequencing (GATC Biotech., Mulhouse, France) and used as templates for in vitro transcription with the MEGAscript Kit (Ambion, Austin, TX, USA) to obtain purified Wn1 and Wn2 RNA. .. RT was carried out in 20-µL volumes with 500 ng of RNA, 1 µg of oligo-dT primers (random primers for bacterial RNA), 1 × ImPromII reaction buffer, 3 mm MgCl2 , 0.5 mm of each dNTP, 20 U of RNasin Ribonuclease Inhibitor and 1 µL of ImProm-II™ (Promega Corp.).

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: Genomic DNA was purified by using the Wizard Genomic DNA purification Kit (Promega). .. The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega).

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin). .. PCR products were purified with a GeneClean III Kit (Bio 101, Carlsbad, California), and cycle-sequenced with Big Dye version 3.1 chemistry following the manufacturer's protocol (PE-ABI).

Article Title: Multiple Signaling Pathways Are Involved in the Interleukine-4 Regulated Expression of DC-SIGN in THP-1 Cell Line
Article Snippet: The mixture of PCR reaction consisted of 0.2 μ L DNA template, 2 μ L forward/reverse primers (10 mM), 4 μ L MgCl2 (25 mM), 4 μ L dNTP (2.5 mM), 5 μ L 10×PCR buffer, 0.5 μ L Pfu DNA polymerase (2 U/μ L, promega), and the final volume was taken to 50 μ L with water. .. The fragments of DC-SIGN promoters were double digested with MLu I and Bgl II and gel purified and ligated into MLu I- and Bgl II-digested pGL-3/Basic and pGL-3/Enhancer luciferase reporter vectors to generate complete DC-SIGN promoter luciferase reporter plasmids and those without AP-1 and Ets-1bingding sites.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Taeyeumjoweetang Affects Body Weight and Obesity-related Genes in Mice
Article Snippet: Paragraph title: Real-Time Reverse Transcription Polymerase Chain Reaction ... The reaction used a reaction solution of total RNA (3–5 μg), oligo d (T) 12–18 (1 μg), 2 μl dNTP (10 mM), MMLV reverse transcriptase (200 U), DTT (10 mM), RNase inhibitor (1 μl; Promega, USA) and a 20 μl buffer solution (50 mM Tris–Cl, pH 8.3, 75 mM KCl, 3 mM MgCl2 ) at 42°C for 60 min, after which cDNA was synthesized.

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: .. The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor. ..

Quantitative RT-PCR:

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: Paragraph title: 4.4. RNA Isolation and RT-qPCR ... One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega).

Nested PCR:

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: .. The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor. ..

Chloramphenicol Acetyltransferase Assay:

Article Title: Genomic Fossils Calibrate the Long-Term Evolution of Hepadnaviruses
Article Snippet: The identity of the different bird species used in this study was verified by sequencing a 420-bp fragment of the mitochondrial NADH dehydrogenase subunit 2 (NADH2) gene ( ) using the following primers: Fwd 5′–AGT CAT TTW GGS AGG AAT CCT G ; Rev 5′–TTC CAY TTC TGA TTY CCA GAA G . .. PCR mix was buffer (5×), 5 µl; MgCl2 (25 mM), 2 µl; dNTP (10 mM), 0.5 µl; primer 1 (10 µM), 1 µl; primer 2 (10 µM), 1 µl; Taq (GoTaq, Promega), 1.25 U; DNA, 30–100 ng; and H2 O up to 25 µl.

Chromatin Immunoprecipitation:

Article Title: GmPHD5 acts as an important regulator for crosstalk between histone H3K4 di-methylation and H3K14 acetylation in response to salinity stress in soybean
Article Snippet: .. ChIP-PCR reactions were set up as follows: 4 ul template (~ < 0.1 nmol) was mixed with 0.4 ul dNTP (10 mM), 0.4 ul forward primer (10 uM), 0.4 ul reverse primer (10 uM), 2 ul 10 × PCR buffer, 0.25 ul Taq polymerase (Promega, Wisconsin, USA), and 1 ul MgCl2 (25 mM). ..

Software:

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: As no homologous porcine EST was identified based on the porcine SAA cDNA sequence, primers F0 et R0 were designed from dog, mink, guinea pig, rabbit and human SAA cDNA sequences, CM59173, in a region relatively well conserved between SAA of different subtypes and different animal origins, using VectorNTI software (Invitrogen, Cergy Pontoise, France). .. PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France).

Article Title: Taeyeumjoweetang Affects Body Weight and Obesity-related Genes in Mice
Article Snippet: The reaction used a reaction solution of total RNA (3–5 μg), oligo d (T) 12–18 (1 μg), 2 μl dNTP (10 mM), MMLV reverse transcriptase (200 U), DTT (10 mM), RNase inhibitor (1 μl; Promega, USA) and a 20 μl buffer solution (50 mM Tris–Cl, pH 8.3, 75 mM KCl, 3 mM MgCl2 ) at 42°C for 60 min, after which cDNA was synthesized. .. The expressed leptin and ghrelin concentrations were calculated relative to the amount of β-actin using LightCycler System software (Roche).

Electrophoresis:

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France). .. Amplification products (10 μl) were loaded onto a 2% TBE agarose gel stained with ethidium bromide (0.01%), subjected to electrophoresis and visualised under a UV transilluminator.

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: RNA integrity was assessed by visualization of 28S and 18S band quality after electrophoresis on a 1% agarose gel. .. One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega).

RNA Extraction:

Article Title: Synthesis of Oxadiazolyl, Pyrazolyl and Thiazolyl Derivatives of Thiophene-2-Carboxamide as Antimicrobial and Anti-HCV Agents
Article Snippet: Paragraph title: RNA Extraction and RT-PCR of HCV RNA ... The thermal cycling protocol was manipulated as follows: 30 min at 42o C for cDNA synthesis followed by 5 min at 95o C and 30 cycles of 1 min at 94°C, 1 min at 55o C and 1 min at 72o C. The nested PCR amplification was performed in 50 µL reaction mixture containing 0.2 mM from each dNTP, 10 µM from each of the reverse nested primer and the forward nested primer, two units of taq DNA polymerase (Promega, Madison, WI, USA), 10 µL from the RT-PCR reaction in a 1X buffer supplied by the Vendor.

Agarose Gel Electrophoresis:

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France). .. Amplification products (10 μl) were loaded onto a 2% TBE agarose gel stained with ethidium bromide (0.01%), subjected to electrophoresis and visualised under a UV transilluminator.

Article Title: No association of the insulin gene VNTR polymorphism with polycystic ovary syndrome in a Han Chinese population
Article Snippet: The PCR reaction mixture contained 50 ng genomic DNA, 0.5 μM of each primer (forward: 5'-AGC AGG TCT GTT CCA AGG-3' and reverse: 5'-CTT GGG TGT GTA GAA GAA GC-3'), 10 mM Tris-HCl (pH 9.0), 50 mM KCl, 1.5 mM MgCl2, 0.1% Triton X-100, 0.2 mM of each dNTP, and 0.75 unit Tag DNA polymerase (Promega, USA) in a final volume of 25 ml. .. The PCR products were 360 bp and were digested with the Hph I restriction enzyme at 37°C for about 18 h. After digested products were electrophoresed on a 2.5% agarose gel and visualized by ethidium bromide staining, the T and A alleles could be distinguished as bands of 231 plus 129 bp and 191 plus 129 bp, respectively.

Article Title: A second generation framework for the analysis of microsatellites in expressed sequence tags and the development of EST-SSR markers for a conifer, Cryptomeria japonica
Article Snippet: 5 ng genomic DNA, 1 × PCR buffer, 200 μM of each dNTP, 1.5 mM MgCl2 , 0.2 μM of each synthesized primer, and 0.25 U of Taq polymerase (Promega Madison, USA), using the following program: 94°C for 5 min, then 40 cycles of 94°C for 30 s, 55–62°C for 30 s and 72°C for 30 s, followed by a final extension at 72°C for 5 min. .. The utility of EST-SSR primers that produced visible bands on the agarose gel was demonstrated by analyzing polymorphisms among 16 individuals of C. japonica from various locations across Japan ( Additional file : Figure S1).

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: RNA integrity was assessed by visualization of 28S and 18S band quality after electrophoresis on a 1% agarose gel. .. One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega).

In Vitro:

Article Title: The biocontrol bacterium Pseudomonas fluorescens Pf29Arp strain affects the pathogenesis-related gene expression of the take-all fungus Gaeumannomyces graminis var. tritici on wheat roots
Article Snippet: These selected amplified sequences, Wn1T7 and Wn2T7 , were verified by DNA sequencing (GATC Biotech., Mulhouse, France) and used as templates for in vitro transcription with the MEGAscript Kit (Ambion, Austin, TX, USA) to obtain purified Wn1 and Wn2 RNA. .. RT was carried out in 20-µL volumes with 500 ng of RNA, 1 µg of oligo-dT primers (random primers for bacterial RNA), 1 × ImPromII reaction buffer, 3 mm MgCl2 , 0.5 mm of each dNTP, 20 U of RNasin Ribonuclease Inhibitor and 1 µL of ImProm-II™ (Promega Corp.).

Ethanol Precipitation:

Article Title: GmPHD5 acts as an important regulator for crosstalk between histone H3K4 di-methylation and H3K14 acetylation in response to salinity stress in soybean
Article Snippet: To reverse the protein-DNA crosslinks, 5 M NaCl was applied to the eluted samples to a final concentration of 200 mM and heated at 65°C for over 4 h. The total DNA was finally recovered from the samples by phenol/chloroform extraction and ethanol precipitation. .. ChIP-PCR reactions were set up as follows: 4 ul template (~ < 0.1 nmol) was mixed with 0.4 ul dNTP (10 mM), 0.4 ul forward primer (10 uM), 0.4 ul reverse primer (10 uM), 2 ul 10 × PCR buffer, 0.25 ul Taq polymerase (Promega, Wisconsin, USA), and 1 ul MgCl2 (25 mM).

Spectrophotometry:

Article Title: Targeted Suppression and Knockout of ASCT2 or LAT1 in Epithelial and Mesenchymal Human Liver Cancer Cells Fail to Inhibit Growth
Article Snippet: A Nanodrop 2000 Micro-Volume UV-Vis spectrophotometer was used to quantify RNA concentrations. .. One µg of total RNA was used from each sample for reverse transcription (RT) reactions, which contained reverse transcriptase, oligo(dT), 5× reaction buffer, MgCl2 , RNasin, dNTP (all from Promega).

Sampling:

Article Title: The identity, distribution, and impacts of non-native apple snails in the continental United States
Article Snippet: Paragraph title: Sampling, DNA extraction, and sequencing ... Reactions were performed in a MJ Research PTC-200 thermal cycler in 50 μl volumes with 1.5 mM MgCl2 , each dNTP at 200 micromolar, 10–100 nanograms of genomic DNA, and 1X Promega buffer B, and 1 U of Taq polymerase (Promega, Madison, Wisconsin).

Produced:

Article Title: Genomic Fossils Calibrate the Long-Term Evolution of Hepadnaviruses
Article Snippet: PCR mix was buffer (5×), 5 µl; MgCl2 (25 mM), 2 µl; dNTP (10 mM), 0.5 µl; primer 1 (10 µM), 1 µl; primer 2 (10 µM), 1 µl; Taq (GoTaq, Promega), 1.25 U; DNA, 30–100 ng; and H2 O up to 25 µl. .. All sequences produced in this study were submitted to GenBank (accession numbers HQ116564–HQ116583).

Article Title: A second generation framework for the analysis of microsatellites in expressed sequence tags and the development of EST-SSR markers for a conifer, Cryptomeria japonica
Article Snippet: 5 ng genomic DNA, 1 × PCR buffer, 200 μM of each dNTP, 1.5 mM MgCl2 , 0.2 μM of each synthesized primer, and 0.25 U of Taq polymerase (Promega Madison, USA), using the following program: 94°C for 5 min, then 40 cycles of 94°C for 30 s, 55–62°C for 30 s and 72°C for 30 s, followed by a final extension at 72°C for 5 min. .. The utility of EST-SSR primers that produced visible bands on the agarose gel was demonstrated by analyzing polymorphisms among 16 individuals of C. japonica from various locations across Japan ( Additional file : Figure S1).

Concentration Assay:

Article Title: GmPHD5 acts as an important regulator for crosstalk between histone H3K4 di-methylation and H3K14 acetylation in response to salinity stress in soybean
Article Snippet: To reverse the protein-DNA crosslinks, 5 M NaCl was applied to the eluted samples to a final concentration of 200 mM and heated at 65°C for over 4 h. The total DNA was finally recovered from the samples by phenol/chloroform extraction and ethanol precipitation. .. ChIP-PCR reactions were set up as follows: 4 ul template (~ < 0.1 nmol) was mixed with 0.4 ul dNTP (10 mM), 0.4 ul forward primer (10 uM), 0.4 ul reverse primer (10 uM), 2 ul 10 × PCR buffer, 0.25 ul Taq polymerase (Promega, Wisconsin, USA), and 1 ul MgCl2 (25 mM).

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: .. The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega). .. The PCR product was purified using GFX PCR DNA and a Gel Band Kit (GE Healthcare), then sequenced using Big Dye v3.1 Kit and 3730 DNA Analyser (Applied Biosystems).

Article Title: Selective stalling of human translation through small-molecule engagement of the ribosome nascent chain
Article Snippet: Translation reactions were transferred to ice and supplemented with 0.5 mM each dNTP, 50 mM Tris-Cl pH 8.3, 9.4 mM MgCl2 , 10 mM DTT, 0.8 U/μL RNasin Plus (Promega), and 0.1 mg/mL cycloheximide. .. The reactions were cooled to room temperature, and 5′-FAM-labelled primers (IDT) were added to a final concentration of 2 μM and AMV reverse transcriptase (Promega) was added to a final concentration of 0.5 U/μL.

DNA Purification:

Article Title: Proteomic Investigation of Aphid Honeydew Reveals an Unexpected Diversity of Proteins
Article Snippet: Genomic DNA was purified by using the Wizard Genomic DNA purification Kit (Promega). .. The PCR mixture contained PCR Buffer, 2 mM MgCl2 , 1 U of Taq polymerase (Fermentas), and dNTP at a concentration of 20 mM (Promega).

Staining:

Article Title: Identification in milk of a serum amyloid A peptide chemoattractant for B lymphoblasts
Article Snippet: PCR was carried out for SAA cDNA amplification, in a total volume of 20 μl containing 100 ng of cDNA, 1 U Taq polymerase (Promega France, Charbonnières, France), 0.2 mM of each dNTP, 0.5 μM of each primers, 1× Taq polymerase buffer (Promega France, Charbonnières, France), 2 mM MgCl2 (Promega France, Charbonnières, France). .. Amplification products (10 μl) were loaded onto a 2% TBE agarose gel stained with ethidium bromide (0.01%), subjected to electrophoresis and visualised under a UV transilluminator.

Article Title: No association of the insulin gene VNTR polymorphism with polycystic ovary syndrome in a Han Chinese population
Article Snippet: The PCR reaction mixture contained 50 ng genomic DNA, 0.5 μM of each primer (forward: 5'-AGC AGG TCT GTT CCA AGG-3' and reverse: 5'-CTT GGG TGT GTA GAA GAA GC-3'), 10 mM Tris-HCl (pH 9.0), 50 mM KCl, 1.5 mM MgCl2, 0.1% Triton X-100, 0.2 mM of each dNTP, and 0.75 unit Tag DNA polymerase (Promega, USA) in a final volume of 25 ml. .. The PCR products were 360 bp and were digested with the Hph I restriction enzyme at 37°C for about 18 h. After digested products were electrophoresed on a 2.5% agarose gel and visualized by ethidium bromide staining, the T and A alleles could be distinguished as bands of 231 plus 129 bp and 191 plus 129 bp, respectively.

Article Title: A second generation framework for the analysis of microsatellites in expressed sequence tags and the development of EST-SSR markers for a conifer, Cryptomeria japonica
Article Snippet: 5 ng genomic DNA, 1 × PCR buffer, 200 μM of each dNTP, 1.5 mM MgCl2 , 0.2 μM of each synthesized primer, and 0.25 U of Taq polymerase (Promega Madison, USA), using the following program: 94°C for 5 min, then 40 cycles of 94°C for 30 s, 55–62°C for 30 s and 72°C for 30 s, followed by a final extension at 72°C for 5 min. .. The PCR products were electrophoretically separated on 2% agarose gels and stained with ethidium bromide to check for successful amplification.

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  • 99
    Promega dntps
    Controlling the extent of pausing by <t>AMV-RT</t> at a 7-EAA site. (A) Sequences of strands employed in primer extension assay, with N indicating the site of the variable nucleotide. (B) Primer extension results for reactions containing 10 μM <t>dNTPs</t> at 42 °C for 45 min. Lanes are labeled as follows for N: −, labeled primer only, no extension; A, adenosine; 7, 7-EAA; B, biotin triazole; and B + S, biotin triazole + monomeric streptavidin. (C) Primer extension results for reactions containing 1 μM dNTPs at 37 °C for 5 min; lanes are labeled the same as in panel B, and the arrow indicates a pause site. (D) Quantification of inhibition of primer extension under the conditions used in panel C; the average for at least three independent primer extension reactions ± standard deviation is plotted.
    Dntps, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 496 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dntps/product/Promega
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    dntps - by Bioz Stars, 2020-02
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    90
    Promega deoxynucleotide triphosphates
    Primer extension with <t>deoxynucleotide</t> triphosphates. The positions of unextended (inverted open triangle), singly extended (inverted black triangle) and doubly extended primers (inverted grey triangle) are indicated. All spectra contain one additional equivalent of primer as a post-reaction standard. (A–D) Incorporation of dGTP for the following primer/template/polymerase combinations. ( A ) P1/LTT/exo – KF. ( B ) SP1/LTT/exo – KF. ( C ) SP1/LTT/exo + KF. ( D ) SP1/LTC/exo + KF. (E–H) Incorporation of dATP for the following primer/template/polymerase combinations. ( E ) P1/LTT/exo – KF. ( F ) SP1/LTT/exo – KF. ( G ) SP1/LTT/exo + KF. ( H ) SP1/LTT/T4 DNAP. All spectra include a signal from primer added post-reaction as an internal standard.
    Deoxynucleotide Triphosphates, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/deoxynucleotide triphosphates/product/Promega
    Average 90 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
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    Image Search Results


    Controlling the extent of pausing by AMV-RT at a 7-EAA site. (A) Sequences of strands employed in primer extension assay, with N indicating the site of the variable nucleotide. (B) Primer extension results for reactions containing 10 μM dNTPs at 42 °C for 45 min. Lanes are labeled as follows for N: −, labeled primer only, no extension; A, adenosine; 7, 7-EAA; B, biotin triazole; and B + S, biotin triazole + monomeric streptavidin. (C) Primer extension results for reactions containing 1 μM dNTPs at 37 °C for 5 min; lanes are labeled the same as in panel B, and the arrow indicates a pause site. (D) Quantification of inhibition of primer extension under the conditions used in panel C; the average for at least three independent primer extension reactions ± standard deviation is plotted.

    Journal: ACS Chemical Biology

    Article Title: Click Modification of RNA at Adenosine: Structure and Reactivity of 7-Ethynyl- and 7-Triazolyl-8-aza-7-deazaadenosine in RNA

    doi: 10.1021/cb500270x

    Figure Lengend Snippet: Controlling the extent of pausing by AMV-RT at a 7-EAA site. (A) Sequences of strands employed in primer extension assay, with N indicating the site of the variable nucleotide. (B) Primer extension results for reactions containing 10 μM dNTPs at 42 °C for 45 min. Lanes are labeled as follows for N: −, labeled primer only, no extension; A, adenosine; 7, 7-EAA; B, biotin triazole; and B + S, biotin triazole + monomeric streptavidin. (C) Primer extension results for reactions containing 1 μM dNTPs at 37 °C for 5 min; lanes are labeled the same as in panel B, and the arrow indicates a pause site. (D) Quantification of inhibition of primer extension under the conditions used in panel C; the average for at least three independent primer extension reactions ± standard deviation is plotted.

    Article Snippet: Samples were then mixed with AMV-RT and dNTP mix so that the final concentrations were as follows: 20 nM RNA, ∼80 nM 32 P-labeled GluR B pre-mRNA 18 nt primer, 10 μM dNTPs, 1× Promega AMV-RT buffer, and 5 units of AMV-RT for the standard extension conditions protocol and 20 nM RNA, ∼80 nM 32 P-labeled GluR B pre-mRNA 18 nt primer, 1 μM dNTPs, 1× Promega AMV-RT buffer, and 5 units of AMV-RT for the low [dNTP] conditions protocol.

    Techniques: Primer Extension Assay, Labeling, Inhibition, Standard Deviation

    In vitro template-primed cDNA synthesis. ( A ) Bordetella bacteriophage DGR diversification of Mtd. mtd contains a variable region ( VR ), which encodes the receptor-binding site of the Mtd protein. Downstream of VR is the template region ( TR ). Adenines in TR (‘A’) are frequently replaced by another base in VR (‘N’). TR is transcribed to produce TR- RNA, which is then reverse transcribed to TR- cDNA. During this process, adenines in TR are mutagenized, as depicted by ‘X’ in TR -cDNA. Adenine-mutagenized TR- cDNA homes to and replaces VR , resulting in diversification of Mtd. bRT is the DGR reverse transcriptase, and avd the DGR accessory variability determinant. ( B ) Sequence elements of the 580 nt DGR RNA template used for reverse transcription reactions. ( C ) bRT-Avd, bRT, or Avd was incubated with the 580 nt DGR RNA and dNTPs, including [α- 32 P]dCTP, for 2h. Products resulting from the incubation were untreated (U), or treated with RNase (+R), DNase (+D), or both RNase and DNase (+R+D), and resolved by 8% denaturing polyacrylamide gel electrophoresis (PAGE). Lane T corresponds to internally-labeled 580 nt DGR RNA as a marker for the size of the template. The positions of the 580 nt band, and 120 and 90 nt cDNA bands are indicated. Nuclease-treated samples were loaded at twice the amount as untreated samples, here and throughout unless otherwise indicated. Lane M here and throughout corresponds to radiolabeled, single-stranded DNA molecular mass markers (nt units). ( D ) DGR RNA templates containing internal truncations in TR . ( E ) Radiolabeled cDNA products resulting from bRT-Avd activity for 2 h with intact (WT) or internally truncated 580 nt DGR RNA as template. Samples were treated with RNase and resolved by denaturing PAGE. The positions of the 120 and 90 nt cDNAs produced from intact template are indicated by red and yellow circles, respectively, as are positions of the correspondingly shorter cDNAs produced from truncated RNA templates. ( F ) Radiolabeled products resulting from bRT-Avd activity for 2 h with the 580 nt DGR RNA as template. Prior to reverse transcription, the RNA template was mock-treated (–Per) or treated with periodate (+Per). Products of the reaction were untreated (U) or treated with RNase (+R), and resolved by 4% (top) or 8% (bottom) denaturing PAGE. In the top gel, the red arrowhead indicates the ∼580 nt species, and the green arrowheads the several ∼540 nt species. In the bottom gel, the black arrowheads indicate the 120 and 90 nt cDNA products. The black vertical line within the gel indicates irrelevant lanes that were removed for display purposes. A 2-fold higher quantity was loaded for +Per samples than –Per samples.

    Journal: Nucleic Acids Research

    Article Title: Template-assisted synthesis of adenine-mutagenized cDNA by a retroelement protein complex

    doi: 10.1093/nar/gky620

    Figure Lengend Snippet: In vitro template-primed cDNA synthesis. ( A ) Bordetella bacteriophage DGR diversification of Mtd. mtd contains a variable region ( VR ), which encodes the receptor-binding site of the Mtd protein. Downstream of VR is the template region ( TR ). Adenines in TR (‘A’) are frequently replaced by another base in VR (‘N’). TR is transcribed to produce TR- RNA, which is then reverse transcribed to TR- cDNA. During this process, adenines in TR are mutagenized, as depicted by ‘X’ in TR -cDNA. Adenine-mutagenized TR- cDNA homes to and replaces VR , resulting in diversification of Mtd. bRT is the DGR reverse transcriptase, and avd the DGR accessory variability determinant. ( B ) Sequence elements of the 580 nt DGR RNA template used for reverse transcription reactions. ( C ) bRT-Avd, bRT, or Avd was incubated with the 580 nt DGR RNA and dNTPs, including [α- 32 P]dCTP, for 2h. Products resulting from the incubation were untreated (U), or treated with RNase (+R), DNase (+D), or both RNase and DNase (+R+D), and resolved by 8% denaturing polyacrylamide gel electrophoresis (PAGE). Lane T corresponds to internally-labeled 580 nt DGR RNA as a marker for the size of the template. The positions of the 580 nt band, and 120 and 90 nt cDNA bands are indicated. Nuclease-treated samples were loaded at twice the amount as untreated samples, here and throughout unless otherwise indicated. Lane M here and throughout corresponds to radiolabeled, single-stranded DNA molecular mass markers (nt units). ( D ) DGR RNA templates containing internal truncations in TR . ( E ) Radiolabeled cDNA products resulting from bRT-Avd activity for 2 h with intact (WT) or internally truncated 580 nt DGR RNA as template. Samples were treated with RNase and resolved by denaturing PAGE. The positions of the 120 and 90 nt cDNAs produced from intact template are indicated by red and yellow circles, respectively, as are positions of the correspondingly shorter cDNAs produced from truncated RNA templates. ( F ) Radiolabeled products resulting from bRT-Avd activity for 2 h with the 580 nt DGR RNA as template. Prior to reverse transcription, the RNA template was mock-treated (–Per) or treated with periodate (+Per). Products of the reaction were untreated (U) or treated with RNase (+R), and resolved by 4% (top) or 8% (bottom) denaturing PAGE. In the top gel, the red arrowhead indicates the ∼580 nt species, and the green arrowheads the several ∼540 nt species. In the bottom gel, the black arrowheads indicate the 120 and 90 nt cDNA products. The black vertical line within the gel indicates irrelevant lanes that were removed for display purposes. A 2-fold higher quantity was loaded for +Per samples than –Per samples.

    Article Snippet: Reactions were carried out in 20 μl containing 1.8 μM bRT-Avd, bRT or Avd, 100 ng/μl RNA template, 100 μM dNTPs (Promega) (or varying concentrations of certain dNTPs), 0.5 μCi/μl [α-32 P]dCTP, 20 units RNase inhibitor (NEB) in 75 mM KCl, 3 mM MgCl2 , 10 mM DTT, 50 mM HEPES, pH 7.5, 10% glycerol for 2 h at 37°C.

    Techniques: In Vitro, Binding Assay, Sequencing, Incubation, Polyacrylamide Gel Electrophoresis, Labeling, Marker, Activity Assay, Produced

    Core DGR RNA. ( A ) Schematic of core DGR RNA. ( B ) Radiolabeled products resulting from bRT-Avd activity for 2 h with the core DGR RNA as template. Prior to the reverse transcription reaction, the RNA template was untreated (-Per) or treated with periodate (+Per). Products from the reaction were untreated (U) or treated with RNase (+R), and resolved by 6% denaturing PAGE. Lane T corresponds to internally-labeled core DGR RNA as a marker for the size of the template. Red arrowheads indicate radiolabeled product bands that migrate at the same position or slower than the core DGR RNA, and green arrowheads ones that migrate faster. The positions of the 120 and 90 nt cDNA bands are indicated. The two panels are from the same gel, with the black line indicating that intermediate lanes were removed. ( C ) Internally-labeled core DGR RNA was not incubated (–), or incubated with bRT-Avd alone or bRT-Avd with 100 μM standard dNTPs (+dNTP), 100 μM dCTP (+CTP), 100 μM dNTPs excluding dCTP (+d(A,T,G)TP), or 100 μM nonhydrolyzeable analog of dCTP (+N-dCTP) for 2 h. Incubation products were resolved by denaturing PAGE. The band corresponding to the 5′ fragment of the cleaved core RNA containing either a deoxycytidine alone (5′+dC) or cDNA (5′+cDNA), and the band corresponding to the 3′ fragment of the RNA are indicated. ( D ) The core DGR RNA was biotinylated at its 3′ end (RNA-Bio), and either reacted with no protein or used as a template for reverse transcription with bRT-Avd. The core DGR RNA in its unbiotinylated form (RNA) was also used as a template for reverse transcription with bRT-Avd. Samples were then purified using streptavidin beads, and the presence of TR -cDNA in the purified samples was assessed by PCR. Products from the PCR reaction were resolved on an agarose gel. ( E ) Radiolabeled products resulting from bRT-Avd activity for 12 h with core, hybrid core dA56, or hybrid core A56 DGR RNA as template. Products were untreated (U) or treated with RNase (+R), and resolved by denaturing PAGE. Separate samples of core dA56 and A56 were 5′ 32 P-labeled for visualization of inputs (I). The positions of the 120 and 90 nt cDNAs are indicated.

    Journal: Nucleic Acids Research

    Article Title: Template-assisted synthesis of adenine-mutagenized cDNA by a retroelement protein complex

    doi: 10.1093/nar/gky620

    Figure Lengend Snippet: Core DGR RNA. ( A ) Schematic of core DGR RNA. ( B ) Radiolabeled products resulting from bRT-Avd activity for 2 h with the core DGR RNA as template. Prior to the reverse transcription reaction, the RNA template was untreated (-Per) or treated with periodate (+Per). Products from the reaction were untreated (U) or treated with RNase (+R), and resolved by 6% denaturing PAGE. Lane T corresponds to internally-labeled core DGR RNA as a marker for the size of the template. Red arrowheads indicate radiolabeled product bands that migrate at the same position or slower than the core DGR RNA, and green arrowheads ones that migrate faster. The positions of the 120 and 90 nt cDNA bands are indicated. The two panels are from the same gel, with the black line indicating that intermediate lanes were removed. ( C ) Internally-labeled core DGR RNA was not incubated (–), or incubated with bRT-Avd alone or bRT-Avd with 100 μM standard dNTPs (+dNTP), 100 μM dCTP (+CTP), 100 μM dNTPs excluding dCTP (+d(A,T,G)TP), or 100 μM nonhydrolyzeable analog of dCTP (+N-dCTP) for 2 h. Incubation products were resolved by denaturing PAGE. The band corresponding to the 5′ fragment of the cleaved core RNA containing either a deoxycytidine alone (5′+dC) or cDNA (5′+cDNA), and the band corresponding to the 3′ fragment of the RNA are indicated. ( D ) The core DGR RNA was biotinylated at its 3′ end (RNA-Bio), and either reacted with no protein or used as a template for reverse transcription with bRT-Avd. The core DGR RNA in its unbiotinylated form (RNA) was also used as a template for reverse transcription with bRT-Avd. Samples were then purified using streptavidin beads, and the presence of TR -cDNA in the purified samples was assessed by PCR. Products from the PCR reaction were resolved on an agarose gel. ( E ) Radiolabeled products resulting from bRT-Avd activity for 12 h with core, hybrid core dA56, or hybrid core A56 DGR RNA as template. Products were untreated (U) or treated with RNase (+R), and resolved by denaturing PAGE. Separate samples of core dA56 and A56 were 5′ 32 P-labeled for visualization of inputs (I). The positions of the 120 and 90 nt cDNAs are indicated.

    Article Snippet: Reactions were carried out in 20 μl containing 1.8 μM bRT-Avd, bRT or Avd, 100 ng/μl RNA template, 100 μM dNTPs (Promega) (or varying concentrations of certain dNTPs), 0.5 μCi/μl [α-32 P]dCTP, 20 units RNase inhibitor (NEB) in 75 mM KCl, 3 mM MgCl2 , 10 mM DTT, 50 mM HEPES, pH 7.5, 10% glycerol for 2 h at 37°C.

    Techniques: Activity Assay, Polyacrylamide Gel Electrophoresis, Labeling, Marker, Incubation, Purification, Polymerase Chain Reaction, Agarose Gel Electrophoresis

    Adenine mutagenesis and template-priming. ( A ) Covalently-linked RNA–cDNA molecule. The linkage is to Sp A56 of the RNA, and the first nucleotide reverse transcribed is TR G117. The RT-PCR product resulting from primers 1 and 2 (blue arrows) is indicated by the dashed red line. ( B ) RT-PCR amplicons from 580 nt DGR RNA reacted with no protein (–), bRT, Avd, or bRT-Avd, separated on a 2% agarose gel and ethidium bromide-stained. The specific amplicon produced from reaction with bRT-Avd shown by the red arrowhead. ( C ) Percentage of substitutions in TR -cDNA determined by sequencing. ( D ) Radiolabeled 120 and 90 nt cDNA products, indicated by arrowheads, resulting from bRT-Avd activity with the 580 nt DGR RNA as template for 2 h (left) or 12 h (right). Either standard dNTPs (dATP, dGTP, dCTP, TTP), as indicated by ‘+’,were present in the reaction, or standard dNTPs excluding dATP (-A), dGTP (–G), or TTP (-T) were present. Products were treated with RNase, and resolved by denaturing PAGE. ( E ) Radiolabeled 120 and 90 nt cDNA products, indicated by arrowheads, resulting from bRT-Avd activity for 2 h with the 580 nt DGR RNA as template with varying TTP (top) or dUTP (bottom) concentrations. Products were treated with RNase, and resolved by denaturing PAGE. ( F ) Radiolabeled 120 and 90 nt cDNA products, indicated by arrowheads, resulting from bRT-Avd activity for 2 h with the 580 nt DGR RNA as template with varying dUTP concentrations. Products were either RNase-treated (top), or both RNase- and UDG-treated (bottom), and resolved by denaturing PAGE.

    Journal: Nucleic Acids Research

    Article Title: Template-assisted synthesis of adenine-mutagenized cDNA by a retroelement protein complex

    doi: 10.1093/nar/gky620

    Figure Lengend Snippet: Adenine mutagenesis and template-priming. ( A ) Covalently-linked RNA–cDNA molecule. The linkage is to Sp A56 of the RNA, and the first nucleotide reverse transcribed is TR G117. The RT-PCR product resulting from primers 1 and 2 (blue arrows) is indicated by the dashed red line. ( B ) RT-PCR amplicons from 580 nt DGR RNA reacted with no protein (–), bRT, Avd, or bRT-Avd, separated on a 2% agarose gel and ethidium bromide-stained. The specific amplicon produced from reaction with bRT-Avd shown by the red arrowhead. ( C ) Percentage of substitutions in TR -cDNA determined by sequencing. ( D ) Radiolabeled 120 and 90 nt cDNA products, indicated by arrowheads, resulting from bRT-Avd activity with the 580 nt DGR RNA as template for 2 h (left) or 12 h (right). Either standard dNTPs (dATP, dGTP, dCTP, TTP), as indicated by ‘+’,were present in the reaction, or standard dNTPs excluding dATP (-A), dGTP (–G), or TTP (-T) were present. Products were treated with RNase, and resolved by denaturing PAGE. ( E ) Radiolabeled 120 and 90 nt cDNA products, indicated by arrowheads, resulting from bRT-Avd activity for 2 h with the 580 nt DGR RNA as template with varying TTP (top) or dUTP (bottom) concentrations. Products were treated with RNase, and resolved by denaturing PAGE. ( F ) Radiolabeled 120 and 90 nt cDNA products, indicated by arrowheads, resulting from bRT-Avd activity for 2 h with the 580 nt DGR RNA as template with varying dUTP concentrations. Products were either RNase-treated (top), or both RNase- and UDG-treated (bottom), and resolved by denaturing PAGE.

    Article Snippet: Reactions were carried out in 20 μl containing 1.8 μM bRT-Avd, bRT or Avd, 100 ng/μl RNA template, 100 μM dNTPs (Promega) (or varying concentrations of certain dNTPs), 0.5 μCi/μl [α-32 P]dCTP, 20 units RNase inhibitor (NEB) in 75 mM KCl, 3 mM MgCl2 , 10 mM DTT, 50 mM HEPES, pH 7.5, 10% glycerol for 2 h at 37°C.

    Techniques: Mutagenesis, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Amplification, Produced, Sequencing, Activity Assay, Polyacrylamide Gel Electrophoresis

    Primer extension with deoxynucleotide triphosphates. The positions of unextended (inverted open triangle), singly extended (inverted black triangle) and doubly extended primers (inverted grey triangle) are indicated. All spectra contain one additional equivalent of primer as a post-reaction standard. (A–D) Incorporation of dGTP for the following primer/template/polymerase combinations. ( A ) P1/LTT/exo – KF. ( B ) SP1/LTT/exo – KF. ( C ) SP1/LTT/exo + KF. ( D ) SP1/LTC/exo + KF. (E–H) Incorporation of dATP for the following primer/template/polymerase combinations. ( E ) P1/LTT/exo – KF. ( F ) SP1/LTT/exo – KF. ( G ) SP1/LTT/exo + KF. ( H ) SP1/LTT/T4 DNAP. All spectra include a signal from primer added post-reaction as an internal standard.

    Journal: Nucleic Acids Research

    Article Title: Single base extension (SBE) with proofreading polymerases and phosphorothioate primers: improved fidelity in single-substrate assays

    doi:

    Figure Lengend Snippet: Primer extension with deoxynucleotide triphosphates. The positions of unextended (inverted open triangle), singly extended (inverted black triangle) and doubly extended primers (inverted grey triangle) are indicated. All spectra contain one additional equivalent of primer as a post-reaction standard. (A–D) Incorporation of dGTP for the following primer/template/polymerase combinations. ( A ) P1/LTT/exo – KF. ( B ) SP1/LTT/exo – KF. ( C ) SP1/LTT/exo + KF. ( D ) SP1/LTC/exo + KF. (E–H) Incorporation of dATP for the following primer/template/polymerase combinations. ( E ) P1/LTT/exo – KF. ( F ) SP1/LTT/exo – KF. ( G ) SP1/LTT/exo + KF. ( H ) SP1/LTT/T4 DNAP. All spectra include a signal from primer added post-reaction as an internal standard.

    Article Snippet: Deoxynucleotide triphosphates (Promega), dideoxynucleotide triphosphates (MBI) and acyclonucleotide triphosphates (NEB) were purchased commercially.

    Techniques: