Fermentas FastDigest Restriction Enzymes ... and FastDigest Green buffers and are able to digest DNA in 5-15 minutes. Peak DNA digestion without star activity is best accomplished with conventional Thermo Scientific restriction enzymes using the Five Buffer System .

3 – Plasmid double-digested with FastDigest EcoRI and KpnI 4 – Plasmid triple-digested with FastDigest EcoRI, KpnI, and SmaI 5 min digestions in FastDigest Green Buffer • Increase the incubation time by 3–5 min if the total reaction volume exceeds 20 μL.

If such a buffer is not indicated, choose the buffer in which both enzymes maintain at least 20% of their activity. ... Alternatively, the optimal buffer can be determined from the chart of common double digestions. Restriction Enzyme Digest Protocol. 1.

4. If such a buffer is not indicated, choose the buffer in which both enzymes Select each enzyme of interest. DoubleDigest conveniently calculates the best enzymatic reaction buffer, enzyme concentrations, incubation conditions, and any additives needed in your double digest reaction. Find great deals on eBay for archie double digest and archie double digest lot. Double Digest Protocol with Standard Restriction Enzymes. Find more protocols and selection guides in the Molecular Biology Guide.

This enables any combination of restriction enzymes to ... Double/multiple restriction digestion Limited due to buffer incompatibility Unlimited - all 176 enzymes are 100% active in … Therefore, enzymes for downstream applications can be directly … 4. FastDigest enzymes can be used to digest plasmid, genomic and viral DNA as well as PCR products and do not show star activity even in prolonged incubations. If the optimal color-coded buffer for both enzymes is the same, use that buffer. Select each enzyme of interest. Double Digestion using Fermentas Color-coded Buffers ™ If the recommended buffer for both enzymes is the same, use that buffer. If not, use the buffer in which both enzymes are most active. As an added convenience, the FastDigest Green Buffer allows for direct loading of the reaction mixture on gels. How do I do a Double Digest?

DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Double Digestion using Fermentas Color-coded Buffers If the recommended buffer for both enzymes is the same, use that buffer. Shop with confidence.

This enables any combination of restriction enzymes to work simultaneously in one reaction tube and eliminates the need for sequential digestions. Air thermostats are not recommended due to slow heat transfer to the reaction mixture. Protocols.io also provides an interactive version of this protocol where you can discover and share optimizations with the research community. 1.

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When using two restriction enzymes at once, first check the enzyme activities in each buffer, using table on of the Restriction Enzyme Buffer Reference. How do I do a Double Digest? If neither buffer is suitable for both REs, refer to the column headed “Y+/Tango™buffer concentration for double digestion”.

digest DNA in 5-15 minutes.

Otherwise, choose an NEBuffer that results in the most activity for both enzymes. Protocol for DNA Digestion with two restriction enzymes. Our restriction enzyme collection has been optimized for digestion using five unique buffers. Use NEBcloner to find the right products and protocols for each in your traditional cloning workflow, including double digestion buffers.

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If not, use the buffer in which both enzymes are most active. It is available for Single-temperature Double Digest, Multi-temperature Double Digest (single buffer), and Sequential Double Digest. If the optimal color-coded buffer for both enzymes is the same, use that buffer. If neither buffer is suitable for both REs, refer to the column headed “Y+/Tango™buffer concentration for double digestion”. Restriction Enzyme Double Digest Buffer Guide. The universal buffer allows rapid single-, double- or multiple DNA digestion within 5-15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps.

and FastDigest Green buffers and are able to digest DNA in 5-15 minutes. Digesting a DNA substrate with two restriction enzymes simultaneously (double digestion) is a common timesaving procedure. Over 215 restriction enzymes are 100% active in CutSmart® Buffer, making double digestion simple. 3.