That application will give you the optimum buffering conditions for both of those enzymes used in the double digest. The application will take into consideration activity of each enzyme in that given buffer, as well as things like star activity.
NEB Interactive Tools. Read Coverage Calculator . Der Double Digest Finder hilft bei der Auswahl des richtigen Reaktionspuffers beim Ansatz von Doppel-Verdaus.
Click here for more information on double digests and a chart of double digests for some common enzyme combinations. To perform a double digest calculation, please select both 1st a Do I need to digest the product directly or to do a purification before digestion and then purify again before after digestion to do the ligation. I am using enzymes from NEB. Thanks. Thermostable Ligase Reaction Temperature Calculator; NEBuilder ® Assembly Tool; Enzyme Finder; Double Digest Finder; NEBcutter ®; REBASE ®; DNA Sequences and Maps Tool; Tm Calculator; PCR Fidelity Estimator; PCR Selector; PolBase ®; NEB Golden Gate Assembly Tool; NEBaseChanger ®; NEBnow Locator; Read Coverage Calculator; Competitor Cross If you choose Restriction Digest, select the 5' and 3' restriction sites you will be using.
Download the Ligation Calculator app for Android Android 2.3 (Gingerbread) Android 2.2 (Froyo) Determine buffer and reaction conditions for experiments requiring two restriction enzymes using the Double Digest Finder. When using either of these tools, look for Time-Saver and HF enzymes for the ultimate in convenience. Use Tm Calculator to calculate optimum annealing temperature for PCR primers when using NEB polymerases and buffers. The HaeIII digest of ΦX174 yields 11 fragments suitable for use as molecular weight standards for agarose gel electrophoresis (1). Comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS. Recommended gel percentage range: 2-3.5%; Optimum separation on 3% ; φX174 DNA-HaeIII Digest visualized by ethidium bromide staining. 1.7% Use this tool to guide your reaction buffer selection when setting up double digests, a common time saving procedure.
Otherwise, choose an NEBuffer that results in the most activity for both enzymes.
Locate commercially available restriction enzymes by category, name, recognition sequence, or overhang.
Otherwise, choose an NEBuffer that results in the most activity for both enzymes. Use NEBcloner to find the right products and protocols for each in your traditional cloning workflow, including double digestion buffers.
Double Digest Finder. Use this tool to guide your reaction buffer selection when setting up double-digests, a common timesaving procedure. Choosing the right buffers will help you to avoid star activity and loss of product.
NEBioCalculator version 1.12.0 That application will give you the optimum buffering conditions for both of those enzymes used in the double digest. The application will take into consideration activity of each enzyme in that given buffer, as well as things like star activity.
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CutSmart® Buffer simplifying double digests; Visit NEBCutSmart.com to learn Online resources, services & software for the restriction analysis of DNA sequences.
Working continuously to be worthy of that distinction, NEB strives to develop enzyme of the highest purity and unparalleled quality. Time-Saver™ qualified for digestion in 5-15 minutes; 100% activity in CutSmart ® Buffer (over 210 enzymes are available in the same buffer) allowing for easier double digests
Double Digest Finder. Use this tool to guide your reaction buffer selection when setting up double-digests, a common timesaving procedure. Choosing the right buffers will help you to avoid star activity and loss of product.
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Comes supplied with 1 vial of Gel Loading Dye, Purple (6X), no SDS. Recommended gel percentage range: 2-3.5%; Optimum separation on 3% ; φX174 DNA-HaeIII Digest visualized by ethidium bromide staining. 1.7% Use this tool to guide your reaction buffer selection when setting up double digests, a common time saving procedure. Choosing the right buffers will help yo You cannot use either one in the double digestion reaction mix, because if you used buffer H in the reaction mix, EcoRI will work with optimal activity (100%) and BamHI will not work (0% activity).
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The minimum videos required to watch are (Digestion with Restriction Enzymes, Standard Protocol for Restriction. Enzyme Digests, and NEB Restriction Enzyme
Add components to a clean tube in the order shown: 1 µL DNA (concentration 1 µg/µL) 2 µL 10x buffer Aug 22, 2014 Explore NEB's entire suite of interactive online tools at www.neb.com/nebtools. NEBioCalculator™.