Learn about our tools that are helping researchers develop diagnostics and vaccines for the SARS-CoV-2 virus. star activity may result from extended digestion. Visit NEBCutSmart.com for further details. This includes personalizing content and advertising. Item# Description List Price Web Price Qty; T1010L: Monarch Plasmid Miniprep Kit - 250 preps: $422.00: $379.80 ADD TO CART: T1010S : Monarch Plasmid Miniprep Kit - 50 preps: $102.00: $91.80 ADD TO CART: T1020L: Monarch DNA Gel Extraction Kit - 250 preps: $594.00: $534.60 ADD TO CART: T1020S: … Enzyme B μl. If star activity is a concern, consider using one of our High Fidelity (HF®) enzymes. The protocol recommends that the first digestion be carried out with the enzyme buffer combination requiring the lowest amount of salt. Set up reaction according to recommended protocol. In general, we recommend 5–10 units of enzyme per µg DNA, and 10–20 units for genomic DNA in a 1 hour digest. Our latest RUO kit, the Luna® SARS-CoV-2 RT-qPCR Multiplex Assay Kit, enables high throughput workflows for real-time detection of SARS-CoV-2 nucleic acid using hydrolysis probes. However, to speed up the screening process, choose one of NEB's enzymes that are Time-Saver qualified. Please note that NEBcloner will also provide detailed double digest protocols using this enzyme. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site. >195 restriction enzymes are Time-Saver qualified, meaning you can digest DNA in 5-15 minutes, or digest DNA safely overnight. All information on the website has been updated to reflect this change. Download a PDF containing pricing for our full product list. international site. Web pricing is applicable only to orders placed online at www.neb.ca. Learn more and request a sample! Use this tool to find the right products and protocols for each step (digestion, end modification, ligation and transformation) of your next traditional cloning experiment. This enzyme has shown to have lower activity on some supercoiled plasmids, with more than 1 unit required to digest 1 μg plasmid DNA. DNA Sequences and Maps Tool. Time-Saver™ qualified for digestion in 5-15 minutes The Destination Plasmid should also have a different antibiotic resistance marker from both the plasmid containing the Upstream Part and the plasmid containing the Downstream Part to avoid … Contact your local subsidiary or distributor. Having supplied restriction enzymes to the research community for over 45 years, NEB has earned the reputation of being the leader in enzyme technologies. This is the Sequential Double Digest Protocol with Standard Restriction Enzymes. To learn more and manage cookies, please refer to our Cookie Statement. It is also used to quickly check the identity of a plasmid by diagnostic digest. ccdB, sacB) in the cloning site to avoid the need for gel purification. These enzymes will digest 1 µg of substrate DNA in 5-15 minutes using 1 µl of enzyme under recommended reaction conditions, and can also be used safely in overnight digestions. DNA Modifying Enzymes & Cloning Technologies, DNA Assembly, Cloning and Mutagenesis Kits, Protein Expression & Purification Technologies, Optimizing Restriction Endonuclease Reactions. Otherwise, choose an NEBuffer that... Set up reaction according to recommended protocol. For cohesive (sticky) ends, incubate at 16°C overnight or room temperature for 10 minutes. Beginning April 2021, we will be gradually transitioning to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. Note that the table... Gently mix the reaction by pipetting up and down and microfuge briefly. Some restriction enzymes require more … X Companion Products. We are excited to announce that we are in the process of switching all reaction buffers to be BSA-free. To save your cart and view previous orders, sign in to your NEB account. Double digests with NEB's restriction enzymes can be set up in CutSmart Buffer. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality. Restriction Digest Protocol. NEB has introduced a line of High-Fidelity (HF®) enzymes that provide added flexibility to reaction setup. Protocol Set up the following reaction in a microcentrifuge tube on ice. A standard reaction is 50 microliters. A specific protocol for single digestion using this restriction enzyme can be accessed using our free online tool, NEBcloner . All of NEB's Restriction enzymes have transitioned to a new buffer system. The desired insert size for the clone library determines which enzymes are selected, as well as the digestion conditions. Kits are available for total RNA purification, plasmid miniprep, gel extraction, and DNA & RNA cleanup. Protocols, Manuals & Usage. Otherwise, choose an NEBuffer that results in the most activity for both enzymes. Restriction Digest Protocol. All information on the website has been updated to reflect this change. Blocked by overlapping dam methylation and CpG methylation. It calls for one microgram of your target DNA, five microliters of the restriction buffer, five to 10 units of enzyme, and then supplementing the rest of the 50 microliters with distilled water. For maximum convenience and value, columns and buffers are also available separately. A specific protocol for single digestion using this restriction enzyme can be accessed using our free online tool, NEBcloner . This is the Double Digest Protocol with Standard Restriction Enzymes, using a common reaction and same incubation temperature for both enzymes. Incubate all three restriction digest reactions at 37°C for 10 minutes and then heat inactivate at 80°C for 20 minutes. Contact your local subsidiary or distributor. Fill out our Technical Support Form, To save your cart and view previous orders, sign in to your NEB account. Protocol for double digestion (20μl system) Pipette the following into a 0.2ml microfuge tube: Enzyme A 1μl . You have been idle for more than 20 minutes, for your security you have been logged out. To learn more and manage cookies, please refer to our Cookie Statement. Setting up a Double Digestion Double digests with NEB's restriction enzymes can be set up in CutSmart Buffer. Beginning April 2021, we will be gradually transitioning to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. The Nucleoside Digestion Mix is a mixture of enzymes that provides a convenient one-step method to generate single nucleosides from DNA or RNA. Please sign back in to continue your session. 1 10 buffer 2μl. For simple, visual assay results, the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit includes a color-changing pH indicator for detection of SARS-CoV-2 nucleic acid amplification. Since CutSmart Buffer includes BSA, there are fewer tubes and pipetting steps to worry about. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality. BsaI-HF ® v2 . Working continuously to be worthy of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality. Restriction Enzyme Digest Protocol: Cutting Close to DNA End EcoRI-HF® We are excited to announce that we are in the process of switching all reaction buffers to be BSA-free. KpnI We are excited to announce that we are in the process of switching all reaction buffers to be BSA-free. Contact our Customer Service Team by Having supplied restriction enzymes to the research community for over 45 years, NEB has earned the reputation of being the leader in enzyme technologies. This includes personalizing content and advertising. RESTRICTION ENZYMES FROM NEB Simplify Reaction Setup and Double Digestion with CutSmart® Buffer Over 210 restriction enzymes are 100% active in a single buffer, CutSmart Buffer, making it significantly easier to set up your double digest reactions. Having supplied restriction enzymes to the research community for over 45 years, NEB has earned the reputation of being the leader in enzyme technologies. Restriction Enzyme Digestion (Protocol for NEB CutSmart® Buffer) Place your order before 7:30pm EST for overnight delivery. Most often, a serial dilution of the selected restriction enzyme(s) is used to digest the starting material and the desired insert size range is isolated by electrophoresis followed by gel extraction of the DNA. Optimized for quantitative analysis by liquid chromatography-mass spectrometry (LC-MS), this reagent eliminates the need for sequential multi-step, time-consuming digestion protocols. Contact your local US Sales Representative. Take advantage of free shipping for any order totaling over $350. Also, find other relevant tools and resources to enable protocol optimization. Learn about our tools that are helping researchers develop diagnostics and vaccines for the SARS-CoV-2 virus. DNA Assembly, Cloning and Mutagenesis Kits, Protein Expression & Purification Technologies, Optimizing Restriction Endonuclease Reactions. Use NEBcloner to find the right products and protocols for each in your traditional cloning workflow, including double digestion buffers. Optimizing Restriction Endonuclease Reactions; Restriction Digest Protocol ; Double Digest Protocol with Standard Restriction Enzymes; Usage Guidelines. Your profile has been mapped to an Institution, please sign back for your profile updates to be completed. Save time and money by placing an order with NEB. Restriction enzyme digestion is commonly used in molecular cloning techniques, such as PCR or restriction cloning. Links to this resource Product Categories: Please note that NEBcloner will also provide detailed double digest protocols using this enzyme. international site. Use NEBcloner to find the right products and protocols for each in your traditional cloning workflow, including double digestion buffers. Beginning April 2021, we will be gradually transitioning to buffers containing Recombinant Albumin (rAlbumin) for restriction enzymes and some DNA modifying enzymes. If your enzyme is a Time-Saver qualified enzyme, it will only require a 5 to 15 minute incubation period. Protocols. Adding products to your cart without being signed in will result in a loss of your cart when you do sign in or leave the site. For simple, visual assay results, the SARS-CoV-2 Rapid Colorimetric LAMP Assay Kit includes a color-changing pH indicator for detection of SARS-CoV-2 nucleic acid amplification. If you don't see your country above, please visit our This method of preparation provides DNA fragments of the desired size with … ddwater rest of the volume . One unit is defined as the amount of enzyme required to digest 1 µg of pBR322 DNA (dammethylated) in 1 hour at 37°C in a total reaction volume of 50 µl. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality. Time-Saver™ qualified for digestion in 5-15 minutes We use cookies to understand how you use our site and to improve the overall user experience. NEB Restriction Enzyme Double Digest Protocol Double digestions can save you time, and this video can offer tips for how to achieve the best results, no matter which of NEB's restriction enzymes you're using. NEBcloner®. Having supplied restriction enzymes to the research community for over 45 years, NEB has earned the reputation of being the leader in enzyme technologies. Your profile has been mapped to an Institution, please sign back for your profile updates to be completed. DNA 0.5-1ug . Time-Saver™ qualified for digestion in 5-15 minutes email or call 1-800-NEB-LABS. For complete digestion of 1 μg of plasmid DNA please follow our recommended digestion protocol. Our latest RUO kit, the Luna® SARS-CoV-2 RT-qPCR Multiplex Assay Kit, enables high throughput workflows for real-time detection of SARS-CoV-2 nucleic acid using hydrolysis probes.

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