Tsap thermosensitive alkaline phosphatase removes 5. Pdf identification and characterization of tsap, a novel. To see articles that cite the use of the dualglo luciferase assay system, visit. And promega 3 employed 2 of these processors in order to. Race, the control rna was consecutively pretreated with thermosensitive alkaline phosphatase tsap, promega, 5. Sap may also be used to treat unincorporated dntps in pcr reactions to prepare. T4 dna ligase is provided with 10x reaction buffer. The dualglo luciferase assay system is designed to allow highthroughput analysis of mammalian cells containing genes for firefly and renilla luciferases, grown in 96 or 384well plates. The dualglo luciferase assay system is designed to work in growth media commonly used for mammalian cells with or without added serum. Reverb and proefx, the latter of which includes choruses, a phaser, a flanger, a couple of delays and so on.
Tsap2, in contrast, was potent against the grampositive bacterium, s. Two peptides, tsap1 and tsap2, from the venom of the. Pdf two peptides, tsap1 and tsap2, from the venom of the. L of icecold wash buffer i by gravity filtration to inactivate tsap. For an overview and information on the development of the dualglo luciferase assay system, see sections 6. The entire miniprep procedure can be completed in 30 minutes or less, depending on the number of samples processed. Active in all promega restriction enzyme buffers, eliminating any cleanup steps or buffer swaps. Panel and bin files for str analysis systems, mass spec quality assurance software, data analysis software.
Description genetic reporter systems are widely used to study eukaryotic gene expression and cellular physiology. Tsap thermosensitive alkaline phosphatase protocol promega. Tsap is active in all promega restriction enzyme buffers, a convenience that allows a single, streamlined restriction enzyme digestiondephosphorylation step. The aminopeptidase ap was found in the proteins produced by t. Alkaline phosphatase, calf intestinal ciap promega. Our product portfolio of over 4,000 products supports a range of life science work in cell biology, dna, rna and protein analysis, drug development, human identification and molecular diagnostics. L of thermosenstive alkaline phosphatase tsap, promega, 2.
Taxonomic identity of landoltia punctata araceae, lemnoideae in. Plasmid dna can be purified in less than 10 minutes, depending on the number of samples processed. The luciferase assay system yields linear results over at least eight orders of magnitude. Tsap thermosensitive alkaline phosphatase, bluewhite cloning qualified. Shrimp alkaline phosphatase rsap is a heat labile alkaline phosphatase purified from a recombinant source. Generally, 100fold greater sensitivity can be achieved over the chloramphenicol acetyltransferase cat assay. Perform everyday lab calculations with the biomath calculators. Please login or register for free to view full text. Cloning the human vitamin d receptor into the ptwin1. In cloning, dephosphorylation prevents relegation of linearized plasmid dna. Tsap1 was of the lowest potency in both assays against all three model test organisms. This system can be used to isolate any plasmid from e.
The reagent is prepared by mixing nanoglo luciferase assay substrate and nanoglo luciferase assay buffer and contains an integral lysis buffer to use directly on cells expressing nanoluc luciferase or culture medium when the luciferase. Instructions for use of products j1250, j1255, j3011, j3015, j4011, j4015. Determining the sequence of rnas is a widely used routine in molecular biology, to which end mostly reverse transcription and consecutive polymerase chain reaction pcr 1 is employed. Tsap is fully active in all promega restriction enzyme reaction buffers, streamlining the. Tsap and sap offer the convenience of heat inactivation. The system provides a rapid method to purify plasmid dna using a silica membrane column.
Tsap thermosensitive alkaline phosphatase catalyzes the removal of 5. The enzyme will not join singlestranded nucleic acids. Rapid ligation is based on the combination of t4 dna ligase with a unique 2x rapid ligation buffer. Pdf two peptides, tsap1 and tsap2, from the venom of. Phosphatase sap promega corporation, catalog number. A new luciferase substrate has enabled promega to create a novel reagent that is more robust and stable when reconstituted, and less aromatic than standard reagents created with luciferin. In this article we test tsap activity in commonly used restriction enzyme buffers from new england biolabs neb. Promega biomath calculators for android apk download. Inactivate sap by heating to 65c for 15 minutes or purify with the wizard sv gel and pcr cleanup. Trichinella spiralis is an intracellular parasite that can cause a serious threat to human health by causing trichinellosis. Dna polymerase i large klenow fragment is a 68kda cterminal fragment of e. Since tsap is active in all promega restriction enzyme buffers, the vector dna can easily be restriction digested and dephosphorylated at the same time. Less than 10 20 moles of luciferase have been detected under optimal conditions.
Tsap thermosensitive alkaline phosphatase promega corporation. Alkaline phosphatase, calf intestinal ciap protocol promega. Structure of yeast argonaute with guide rna nature. As a general guideline, for reactions containing up to 1g of dna, add 15 units of restriction enzyme and the amount of tsap listed below to the vector dna in a total reaction volume of 20. Tsap thermosensitive alkaline phosphatase is active in all promega restriction enzyme buffers, making it an ideal choice for convenient, rapid dephosphorylation of dna. One mbu is defined as the amount of enzyme required to dephosphorylate 1. Nanoglo luciferase assay system provides a simple, singleaddition reagent that generates a glowtype signal in the presence of nanoluc luciferase. M0290 or shrimp alkaline phosphatase sap promega corporation, catalog number. Promega corporation is a worldwide leader in applying biochemistry and molecular biology to the development of innovative, highvalue products for the life sciences.
Tsap thermosensitive alkaline phosphatase activity. Promega manufacturing and delivery systems continue to be fully. Activity, sdspagepurity, endonuclease, dnase, rnase, bluewhite cloning assay. Enzymes were removed from the reaction using pcr cleanup bay gene, inc. To be the most responsive supplier of biological reagents and reagent systems used in research and applied technology applications worldwide.
Brightglo luciferase assay system protocol promega. Molecular characterization of trichinella spiralis. Download sds pdf 67 kb english united states certificate of. Simple and inexpensive threestep rapid amplification of cdna 5. To inactivate tsap, the reaction was quenched with 1. Simple and inexpensive threestep rapid amplification of cdna. Downloadable software and firmware for promega instruments, including luminometers, fluorometers and dna purification instruments. Com number 95 january 2007 introduction tsap, thermosensitive alkaline phosphatase cat. Our teams are in regular contact with suppliers and distributors worldwide and are taking all steps necessary to address both demands for diagnostic tools and reliable delivery of all products as quickly as possible. The robust phosphatase activity effectively dephosphorylates all dna termini 5. Shrimp alkaline phosphatase sap thermo fisher scientific.
The description of promega biomath calculators perform everyday lab calculations with a single app. Instructions for use of products e2920, e2940, e2980. Tsap thermosensitive alkaline phosphatase, 100 units, promega. Rna was ligated to preadenylated adaptor dna in a 50. The pureyield plasmid miniprep system isolates highquality plasmid dna for use in eukaryotic transfection and in vitro expression experiments. Dna dephosphorylation under the appropriate con ditions. In press, journal preproof what are journal preproof articles. A new thermosensitive alkaline phosphatase promega. Dualluciferase reporter assay system protocol promega.
Purchase the subcloning tools bundle, and get ligafast rapid dna ligation system, tsap thermosensitive alkaline phosphatase, benchtop 100bp dna ladder, wizard sv gel and pcr cleanup system and pureyield plasmid miniprep system for one low price. Tsap is active in all promega restriction enzyme buffers, including multicore buffer, as well as both gotaq buffers. Pureyield plasmid miniprep system protocol promega. Tsap a new thermosensitive alkaline phosphatase promega.
The ptwin1 was also treated with thermal sensitive alkaline phosphatase, tsap promega, for an additional 15 min at 37 c. T4 dna ligase catalyzes the joining of two cohesive or bluntended strands of dna between the 5. Only tsap does not require a special buffer and is active in all promega restriction enzyme buffers, making it the most convenient and costeffective choice. The pd 1pd l1 blockade bioassay is a bioluminescent cellbased assay that can be used to measure the potency and stability of antibodies and other biologics designed to block the pd1pdl1 interaction. Active in promega restriction buffers, tsap will allow simultaneous restriction enzyme digestion and. Promega corporation is a leader in providing innovative solutions and technical support to the life sciences industry. The oneglo luciferase assay system employs new assay chemistry to provide a robust, homogeneous assay. Simple and inexpensive threestep rapid amplification of. Promega corporation 2800 woods hollow road madison, wi 537115399 usa toll free in usa 8003569526 6082744330 fax 6082772516 tm033 revised 615 5 3. The ligafast rapid dna ligation system is designed for the efficient ligation of stickyended dna inserts into plasmid vectors in just 5 minutes bluntended inserts in as little as 15 minutes. The human sex hormone binding globulin shbg, testisspecific gene with ankyrin repeats and pest domain tsap, and patched domain containing 2 ptchd2 were identified by in silico cloning 8. The biomath calculators provide a range of functions essential to molecular biology experiments, including nucleic acid and protein conversions, melting temperature, temperature conversion, molarity and dilution calculations. Purchase the subcloning tools bundle, and receive the ligafast rapid dna ligation system, tsap thermosensitive alkaline phosphatase, benchtop 100bp dna ladder, wizard sv gel and pcr cleanup system and pureyield plasmid miniprep system for one low price. Here we describe tsap, a new thermosensitive alkaline phosphatase.