In this video, you will learn how double digests are made even easier with a tool, NEBCloner. NEBCloner provides a nice protocol taking into account buffer, temperature, additive, as well as star activity differences. Let's see how. Open up NEBCloner and select digestion. Next, choose the two enzymes that you would like to digest simultaneously.

240 County Road Ipswich, MA 01938-2723 978-927-5054 (Toll Free) 1-800-632-5227 Fax: 978-921-1350 Info@neb.com

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(NEB) as follows: a 5.24 kb fragment obtained by an Eco RV- Spe I-double digest of AH452 (pXL- BacII-FRT_3xP3DsRed_FRT3_loxN-PUbAmCyan-lox2272 )

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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; 100% activity in rCutSmart ™ Buffer (over 215 enzymes are available in the same buffer) simplifying double digests

This is the Double Digest Protocol with Standard Restriction Enzymes, using a common reaction and same incubation temperature for both enzymes. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality. Methylation-sensitive restriction enzyme; Time-Saver™ qualified for digestion in 5-15 minutes; 100% activity in rCutSmart ™ Buffer (over 215 enzymes are available in the same buffer) simplifying double digests This is the Sequential Double Digest Protocol with Standard Restriction Enzymes. If there is no buffer in which the two enzymes exhibit > 50% activity, this sequential digest DoubleDigest conveniently calculates the best enzymatic reaction buffer, enzyme concentrations, incubation conditions, and any additives needed in your double digest reaction. Peak DNA digestion without star activity is best accomplished with conventional Thermo Scientific restriction enzymes using the Five Buffer System . DoubleDigest conveniently calculates the best enzymatic reaction buffer, enzyme concentrations, incubation conditions, and any additives needed in your double digest reaction. Peak DNA digestion without star activity is best accomplished with conventional Thermo Scientific restriction enzymes using the Five Buffer System .

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Double Digest Finder. Use this tool to guide your reaction buffer selection when setting up double-digests, a common timesaving procedure.

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is a 10X buffer supplied with over 215 restriction enzymes, simplifying double digests. NEB #, Component Name, Component #, Stored at (°C), Amount

DoubleDigest conveniently calculates the best enzymatic reaction buffer, enzyme concentrations, incubation conditions, and any additives needed in your double digest reaction. Peak DNA digestion without star activity is best accomplished with conventional We recommend using the Double Digest Finder and use the buffer it recommends. For example, you have old buffer NEBuffer 1 with one of the enzymes and CutSmart Buffer with the second enzyme. If Double Digest Finder recommends to use NEBuffer 1.1, then you can use NEBuffer 1 + BSA for the double digest.

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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.

Choosing the right buffers will help you to avoid star activity and loss of product. A. We recommend using the Double Digest Finder and use the buffer it recommends. For example, you have old buffer NEBuffer 1 with one of the enzymes and CutSmart Buffer with the second enzyme. If Double Digest Finder recommends to use NEBuffer 1.1, then you can use NEBuffer 1 + BSA for the double digest. Restriction Digest Protocol A specific protocol for single digestion using this restriction enzyme can be accessed using our free online tool, NEBcloner.Please note that NEBcloner will also provide detailed double digest protocols using this enzyme. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled quality. 100% activity in CutSmart ® Buffer (over 215 enzymes are available in the same buffer) simplifying double digests Supplied with 1 vial of Gel Loading Dye, Purple (6X) DoubleDigest conveniently calculates the best enzymatic reaction buffer, enzyme concentrations, incubation conditions, and any additives needed in your double digest reaction.

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100% activity in CutSmart ® Buffer (over 215 enzymes are available in the same buffer) simplifying double digests Supplied with 1 vial of Gel Loading Dye, Purple (6X) DoubleDigest conveniently calculates the best enzymatic reaction buffer, enzyme concentrations, incubation conditions, and any additives needed in your double digest reaction.