The 10X FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading. The blue dye migrates with 3–5 kb DNA fragments in a 1% agarose gel and has an excitation peak of 424 nm. The yellow dye migrates faster than 10 bp DNA fragments in a 1% agarose gel and has an excitation peak of 615 nm.
What are thermo enzymes?
Thermo Scientific FastDigest restriction enzymes are an advanced line of enzymes which offer: Complete digestion in 5-15 minutes. 100% buffer compatibility with downstream applications. Direct loading on gels.
How do you do restriction digestion?
Restriction Enzyme Digest Protocol
- Add components to a clean tube in the order shown:
- Incubate the reaction at digestion temperature (usually 37 °C) for 1 hour.
- Stop the digestion by heat inactivation (65 °C for 15 minutes) or addition of 10 mM final concentration EDTA.
Do Neb enzymes work in fast digest buffer?
I have used a combination of NEB and FastDigest Enzymes: it seems to work fine for me, in fact well I would say. Make sure to use FastDigest and HF enzymes of each, but use 2ul of HF enzyme rather then one, and increase slightly the concentration of the buffer, so like 1.1X. DNA and Water to 40ul.
What is FD buffer?
The FastDigest Green Buffer and Thermo Scientific FastDigest Buffer are proprietary digestion buffers which support 100% activity of all FastDigest restriction enzymes. This system allows for double and multiple digestions with any combination of enzymes. No sequential digestions or buffer changes are needed.
Which enzymes digest fast?
DNA/RNA modifying enzymes, such as ligases, phosphatases, kinases and mesophilic DNA polymerases have 100% activity in FastDigest and FastDigest Green Buffer.
Are enzymes Thermolabile?
Thermolabile refers to a substance which is subject to destruction, decomposition, or change in response to heat. This term is often used to describe biochemical substances. Enzymes are also thermolabile and lose their activity when the temperature rises.
What is the purpose of the buffer in a restriction enzyme reaction?
Major function of the buffer is to maintain pH of the reaction (usually, 8.0) and provide a favorable environment for the enzyme to function.
What are the components of a restriction digest?
The components of a typical restriction digestion reaction include the DNA template, the restriction enzyme of choice, a buffer and sometimes BSA protein. The reaction is incubated at a specific temperature required for optimal activity of the restriction enzyme and terminated by heat.
What is a digestion buffer?
Differex™ Digestion Buffer is used in differential extractions of samples that contain a mixture of epithelial and sperm cells. A digestion combination of this buffer and Proteinase K can be used to selectively lyse epithelial cells while leaving sperm cells intact.
Is the Fermentas FastDigest buffer suitable for double digestions?
Initially, Fermentas stated on their web pages, that “The FastDigest™ buffer is suitable for double digestions with both FastDigest™ enzymes and regular restriction enzymes” ( March 15, 2006; according to the Internet Archive).
What is the 10x FastDigest buffer?
The 10X FastDigest Buffer is a proprietary digestion buffer which supports 100% activity of all FastDigest restriction enzymes. DNA/RNA modifying enzymes, such as ligases, phosphatases, kinases and mesophilic DNA polymerases also have 100% activity in FastDigest Buffer. Benefits:
Why choose Thermo Scientific Fermentas FastDigest?
Our FastDigest line of restriction enzymes is ideal for use in applications that require high purity reaction components, performance reliability and simple reaction set-up. Thermo Scientific Fermentas FastDigest enzymes are an advanced line of restriction enzymes for rapid DNA digestion.
How do I add DNA/RNA modifying enzymes to FastDigest green buffer?
DNA/RNA modifying enzymes, such as ligases, phosphatases, kinases and mesophilic DNA polymerases have 100% activity in FastDigest and FastDigest Green Buffer. Therefore, enzymes used in downstream applications can be directly added to the FastDigest reaction mix. No buffer changes or purification steps are needed.
