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Triphosphate
Triphosphate
Triphosphate is a chemical compound composed of three phosphate groups covalently linked together.
It serves as an important energy-carrying molecule in many biological processes, such as cellular respiration and DNA/RNA synthesis.
Triphospahtes play a crucial role in the activation and transfer of phosphate groups, making them essential for various metabolic pathways and signaling cascades within living organisms.
Triphosphates are found in a variety of biomolecules, including adenosine triphosphate (ATP), guanosine triphosphate (GTP), and cytidine triphosphate (CTP), among others.
Understanding the structure, function, and regulation of triphosphates is crucial for the study of cellular energetics and the development of therapeutic interventions targeting these vital biochemical compounds.
It serves as an important energy-carrying molecule in many biological processes, such as cellular respiration and DNA/RNA synthesis.
Triphospahtes play a crucial role in the activation and transfer of phosphate groups, making them essential for various metabolic pathways and signaling cascades within living organisms.
Triphosphates are found in a variety of biomolecules, including adenosine triphosphate (ATP), guanosine triphosphate (GTP), and cytidine triphosphate (CTP), among others.
Understanding the structure, function, and regulation of triphosphates is crucial for the study of cellular energetics and the development of therapeutic interventions targeting these vital biochemical compounds.
Most cited protocols related to «Triphosphate»
BLOOD
DNA Replication
Erythrocytes
Factor VII
Pharmaceutical Preparations
Phosphoric Monoester Hydrolases
Therapeutics
triphosphate
Volumes, Packed Erythrocyte
mRNAs were transcribed from linearized plasmids encoding firefly Luc, codon-optimized mEPO, EGFP, or hiNOS using T7 RNA polymerase (MEGAscript T7 Transcription Kit, Thermo Fisher Scientific, Darmstadt, Germany). The mRNAs were transcribed to contain the 5′ UTR derived from the tobacco etch virus 5′ leader RNA (TEV).34 (link) Further, a 100-nt-long poly(A) tail interrupted by a short linker (A30LA70) was transcribed from the corresponding DNA templates. For the generation of nucleoside-modified mRNAs, uridine 5’-triphosphate (UTP) was replaced with the triphosphate derivative of m1Ψ (m1ΨTP) in the transcription reaction. Capping of the IVT mRNAs was performed co-transcriptionally using the trinucleotide cap1 analog CleanCap (TriLink, San Diego, CA, USA). After DNase digestion, the synthesized mRNA was isolated from the reaction mix by precipitation with half volume of 8 M LiCl solution (Sigma-Aldrich, Hamburg, Germany), and finally the pellet was dissolved in nuclease-free water. The RNA concentration was determined using a Nanodrop 2000c spectrophotometer (Thermo Fisher Scientific). Aliquots of denatured IVT mRNAs were analyzed by electrophoresis in non-denaturing 1.4% agarose gels containing 0.005% (v/v) GelRed nucleic acid gel stain.35 (link) RiboRuler High Range RNA Ladder (Thermo Fisher Scientific) was loaded as a molecular weight marker.
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5' Untranslated Regions
bacteriophage T7 RNA polymerase
Codon
Deoxyribonucleases
Digestion
Electrophoresis
Fireflies
Nucleic Acids
Nucleosides
Plasmids
Poly(A) Tail
RNA, Messenger
Sepharose
Stains
Tobacco etch virus
Transcription, Genetic
triphosphate
Uridine Triphosphate
The V1–V2 region of the 16S rRNA gene was amplified using forward primer (5′-CCATCTCATCCCTGCGTGTCTCCGACTCAGNNNNNNNNNNagrgtttgatymtggctcag-3′) containing the 454 primer A, a unique 10-bp barcode sequence for each sample (indicated in N), and 27Fmod (5′-agrgtttgatymtggctcag) in which the third base A in the original primer 27F was changed to R, and reverse primer (5′-CCTATCCCCTGTGTGCCTTGGCAGTCTCAGtgctgcctcccgtaggagt-3′) containing the 454 primer B and reverse primer 338R (5′-tgctgcctcccgtaggagt). PCR was performed in 1 × Ex Taq PCR buffer (50 µl), deoxynucleoside triphosphate (2.5 mM), Ex Taq polymerase (Takara Bio, Inc., Shiga), each primer (10 μM), and 40 ng of extracted DNA under conditions of 2 min at 96°C, 20 cycles of 96°C for 30 s, 55°C for 45 s, and 72°C for 1 min, and a final extension of 72°C for 10 min on a 9700 PCR system (Life Technologies Japan, Ltd, Tokyo, Japan). PCR products of approximately 370 bp were confirmed by agarose gel electrophoresis, purified by AMPure XP magnetic purification beads (Beckman Coulter, Inc., Brea, CA, USA), and quantified using the Quant-iT PicoGreen dsDNA Assay Kit (Life Technologies Japan, Ltd, Tokyo, Japan). Mixed samples were prepared by pooling approximately equal amounts of PCR amplicons from each sample and subjected to 454 GS FLX Titanium or 454 GS JUNIOR (Roche Applied Science) sequencing according to the manufacturer's instructions.
Biological Assay
Buffers
DNA, Double-Stranded
Electrophoresis, Agar Gel
Genes
Oligonucleotide Primers
PicoGreen
RNA, Ribosomal, 16S
Taq Polymerase
Titanium
triphosphate
Brucella DNA was prepared as previously described [27 (link)]. PCR amplification was performed in a total volume of 15 μl containing 1ng of DNA, 1× PCR Reaction Buffer, 1 U of Taq DNA polymerase (Qbiogen, Illkirch, France), 200 μM of each deoxynucleotide triphosphate, and 0.3 μM of each flanking primer as described previously [15 (link)].
Amplifications were performed in a MJ Research PTC200 thermocycler. An initial denaturation step at 96°C for 5 minutes was followed by 30 cycles of denaturation at 96°C for 30 s, primer annealing at 60°C for 30 s, and elongation at 70°C for 1 min. The final extension step was performed at 70°C for 5 min.
Two to five microliters of the amplification product were loaded on a 3% standard agarose gel for analyzing tandem repeats with a unit length shorter than 10 bp and on a 2% standard agarose gel for all others, and run under a voltage of 8 V/cm until the bromophenol blue dye had reached the 20 cm position. Gels were stained with ethidium bromide, visualized under UV light, and photographed (Vilber Lourmat, Marnes-la-Vallée, France). A 100-bp and a 20-bp ladder (EZ Load 100 pb or 20 bp PCR Molecular Ruler, Biorad, Marnes-la-Coquette, France) were used as molecular size markers depending on the tandem repeat unit length. Gel images were managed using the Bionumerics software package (version 4.0, Applied-Maths, Belgium).
Amplifications were performed in a MJ Research PTC200 thermocycler. An initial denaturation step at 96°C for 5 minutes was followed by 30 cycles of denaturation at 96°C for 30 s, primer annealing at 60°C for 30 s, and elongation at 70°C for 1 min. The final extension step was performed at 70°C for 5 min.
Two to five microliters of the amplification product were loaded on a 3% standard agarose gel for analyzing tandem repeats with a unit length shorter than 10 bp and on a 2% standard agarose gel for all others, and run under a voltage of 8 V/cm until the bromophenol blue dye had reached the 20 cm position. Gels were stained with ethidium bromide, visualized under UV light, and photographed (Vilber Lourmat, Marnes-la-Vallée, France). A 100-bp and a 20-bp ladder (EZ Load 100 pb or 20 bp PCR Molecular Ruler, Biorad, Marnes-la-Coquette, France) were used as molecular size markers depending on the tandem repeat unit length. Gel images were managed using the Bionumerics software package (version 4.0, Applied-Maths, Belgium).
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Biological Markers
Bromphenol Blue
Brucella
Buffers
Ethidium Bromide
Oligonucleotide Primers
Sepharose
Tandem Repeat Sequences
Taq Polymerase
triphosphate
Ultraviolet Rays
To amplify the TPI fragment from various Giardia isolates, a nested PCR protocol was developed that used primers complementary to the conserved published TPI nucleotide sequences of various Giardia parasites downloaded from GenBank: G. duodenalis (U57897, AF06957 to AF069563, L02116, L02120), G. muris (AF069565), and G. ardeae (AF069564). For the primary PCR, a PCR product of 605 bp was amplified by using primers AL3543 [5′-AAATIATGCCTGCTCGTCG-3′] and AL3546 [5′-CAAACCTTITCCGCAAACC-3′]. The PCR reaction comprised 0.25–2.0 μL of DNA, 200 μM each of deoxynucleoside triphosphate (dNTP), 1X PCR buffer (Perkin Elmer, Wellesley, MA), 3.0 mM MgCl2, 5.0 U of Taq polymerase (GIBCO BRL, Frederick, MD), and 200 nM of each primer in a total of 100-μL reaction. The reactions were performed for 35 cycles (94°C for 45 s, 50°C for 45 s, and 72°C for 60 s) in a Perkin-Elmer GeneAmp PCR 9700 thermocycler, with an initial hot start (94°C for 5 min) and a final extension (72°C for 10 min). For the secondary PCR, a fragment of 530 bp was amplified by using 2.5 μL of primary PCR reaction and primers AL3544 [5′-CCCTTCATCGGIGGTAACTT-3′] and AL3545 [5′-GTGGCCACCACICCCGTGCC-3′]. The conditions for the secondary PCR were identical to the primary PCR. The PCR products were analyzed by agarose gel electrophoresis and visualized after ethidium bromide staining.
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Buffers
Conserved Sequence
Electrophoresis, Agar Gel
Ethidium Bromide
Giardia
Magnesium Chloride
Nested Polymerase Chain Reaction
Oligonucleotide Primers
Parasites
Taq Polymerase
triphosphate
Most recents protocols related to «Triphosphate»
For the adenosine triphosphate (ATP) assay, approximately 3000 cells were seeded in each well of a 96‐well plate overnight. Subsequently, the cells were treated with either 1 mm sodium phosphate (pH 6.8) or 1 mm polyP for the indicated durations. The change in cellular ATP content in each well was measured using the CellTiter‐Glo® 2.0 Assay kit (Promega Corporation). Chemical luminescence was detected using the AVRO MX instrument (PerkinElmer Inc., Waltham, MA, USA).
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The adenosine triphosphate (ATP) content in sperm was measured using an ATP assay kit (EZScreenTM ATP Colorimetric Assay Kit, Biovision, USA). Briefly, sperm samples (3×106 cells/mL) were homogenized with assay buffer (100 μL). Samples were treated with ATP reaction mixture in 96-well plates and gently shaken on a shaker for 2 min to induce sperm lysis. Samples were incubated in the dark for 30 min at room temperature. Finally, absorbance was measured at 570 nm using an enzyme marker (Tecan Infinite M200, Switzerland).
To quantify the metabolic activity of Vorganoids formed at different stages. RIPA buffer was added to lyse the Vorganoids, which were subsequently centrifuged at 4 °C for 5 min at 12000 rpm, after which the supernatant was removed for adenosine triphosphate (ATP) determination. The ATP assay working solution was prepared according to the kit instructions (S0026, Beyotime, China), and an ATP standard curve was constructed. The ATP working solution (100 μL) was added to the test wells and allowed to stand for 3-5 min at room temperature to eliminate background effects. Then, an appropriate amount of sample or standard was added and mixed well. The relative light unit value was measured using a chemiluminescence metre.
The intracellular adenosine triphosphate (ATP) content was quantified using an Enhanced ATP Assay Kit (Beyotime, Nanjing, China) following the manufacturer’s instructions. Briefly, cells were seeded in 6-well plates at a density of 2 × 105 cells/well and cultured overnight. Subsequently, the cells were exposed to 8 Gy radiation and treated with MOTS-c at a concentration of 10 μM for 24 h. Lastly, cell lysis was performed using a lysis reagent followed by centrifugation at 12,000× g for 5 min at 4 °C. The supernatant was then collected to measure ATP levels.
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Adenosine triphosphate (ATP) content was determined by using an enhanced ATP assay kit (Beyotime Biotechnology) according to the manufacturer’s instructions. H9c2 cells and myocardial tissue were lysed with ATP lysis buffer and centrifuged at 10,000 g for 10 min at 4°C. The supernatants were collected and stored on ice. Before the ATP test, 100 μl of ATP working solution was added to 1.5 ml eppendor tubes and incubated for 5 min at room temperature. Next, the supernatant was transferred to 100 μl of ATP working solution and the amount of luminescence emitted was measured immediately with a luminometer.
Top products related to «Triphosphate»
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ATP is a laboratory instrument used to measure the presence and concentration of adenosine triphosphate (ATP) in various samples. ATP is a key molecule involved in energy transfer within living cells. The ATP product provides a reliable and accurate method for quantifying ATP levels, which is useful in applications such as microbial detection, cell viability assessment, and ATP-based assays.
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The In Situ Cell Death Detection Kit is a laboratory product designed for the detection of programmed cell death, or apoptosis, in cell samples. The kit utilizes a terminal deoxynucleotidyl transferase (TdT) to label DNA strand breaks, allowing for the visualization and quantification of cell death. The core function of this product is to provide researchers with a tool to study and analyze cell death processes.
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The ATP assay kit is a tool used to measure the levels of adenosine triphosphate (ATP) in biological samples. ATP is a crucial energy-carrying molecule found in all living cells. The kit provides a method to quantify ATP concentrations, which can provide insights into cellular metabolism and energy production.
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TRIzol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components designed for the isolation of total RNA, DNA, and proteins from a variety of biological samples. The reagent maintains the integrity of the RNA while disrupting cells and dissolving cell components.
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The CellTiter-Glo Luminescent Cell Viability Assay is a quantitative method for determining the number of viable cells in a cell-based assay. The assay measures the amount of ATP present, which is an indicator of metabolically active cells.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
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Taq DNA polymerase is a thermostable enzyme used for DNA amplification in Polymerase Chain Reaction (PCR) applications. It is isolated from the thermophilic bacterium Thermus aquaticus, and its core function is to catalyze the synthesis of new DNA strands complementary to a template DNA sequence.
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Adenosine 5'-triphosphate (ATP) is a naturally occurring nucleotide that serves as the primary energy currency in living cells. It is composed of an adenine base, a ribose sugar, and three phosphate groups. ATP plays a crucial role in various cellular processes, including energy storage and transfer, signal transduction, and the regulation of metabolic pathways.
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Bovine serum albumin (BSA) is a common laboratory reagent derived from bovine blood plasma. It is a protein that serves as a stabilizer and blocking agent in various biochemical and immunological applications. BSA is widely used to maintain the activity and solubility of enzymes, proteins, and other biomolecules in experimental settings.
More about "Triphosphate"
Triphosphates are a class of chemical compounds composed of three covalently linked phosphate groups, known for their crucial role in various biological processes.
These energy-rich molecules, such as adenosine triphosphate (ATP), guanosine triphosphate (GTP), and cytidine triphosphate (CTP), are essential for cellular respiration, DNA/RNA synthesis, and a wide range of metabolic pathways.
Triphosphates serve as the primary energy currency in living organisms, powering a multitude of cellular functions.
They are involved in the activation and transfer of phosphate groups, making them indispensable for signaling cascades, enzymatic reactions, and the regulation of various biochemical processes.
The structure and dynamics of triphosphates are closely linked to their ability to store and release energy, which is crucial for the maintenance of cellular homeostasis and the efficient functioning of living systems.
In the context of biological research, triphosphates are often studied using techniques such as the ATP assay kit, which measures the levels of ATP, and the CellTiter-Glo Luminescent Cell Viability Assay, which utilizes ATP as an indicator of cell viability.
Additionally, the TRIzol reagent, commonly used for RNA extraction, relies on the properties of triphosphates to facilitate the isolation and purification of nucleic acids.
Undestanding the structure, function, and regulation of triphosphates is essential for advancing our knowledge of cellular energetics and metabolic processes.
This knowledge can inform the development of therapeutic interventions targeting these vital biochemical compounds, potentially leading to breakthroughs in the treatment of various diseases and the optimization of research workflows.
These energy-rich molecules, such as adenosine triphosphate (ATP), guanosine triphosphate (GTP), and cytidine triphosphate (CTP), are essential for cellular respiration, DNA/RNA synthesis, and a wide range of metabolic pathways.
Triphosphates serve as the primary energy currency in living organisms, powering a multitude of cellular functions.
They are involved in the activation and transfer of phosphate groups, making them indispensable for signaling cascades, enzymatic reactions, and the regulation of various biochemical processes.
The structure and dynamics of triphosphates are closely linked to their ability to store and release energy, which is crucial for the maintenance of cellular homeostasis and the efficient functioning of living systems.
In the context of biological research, triphosphates are often studied using techniques such as the ATP assay kit, which measures the levels of ATP, and the CellTiter-Glo Luminescent Cell Viability Assay, which utilizes ATP as an indicator of cell viability.
Additionally, the TRIzol reagent, commonly used for RNA extraction, relies on the properties of triphosphates to facilitate the isolation and purification of nucleic acids.
Undestanding the structure, function, and regulation of triphosphates is essential for advancing our knowledge of cellular energetics and metabolic processes.
This knowledge can inform the development of therapeutic interventions targeting these vital biochemical compounds, potentially leading to breakthroughs in the treatment of various diseases and the optimization of research workflows.