How do restriction enzymes help digestion?
Protocol for DNA Digestion with a Single Restriction Enzyme 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. The digested DNA is ready for use in research applications.
What is the purpose of a double restriction digest?
A double digest is one where two restriction enzymes are used to digest DNA in a single reaction. In this case you will be using EcoR I and BamH I. There is only one site in the plasmid vector for each of these enzymes and they are located on either side of your insert DNA.
What is gel electrophoresis used for?
Gel electrophoresis is a laboratory method used to separate mixtures of DNA, RNA, or proteins according to molecular size. In gel electrophoresis, the molecules to be separated are pushed by an electrical field through a gel that contains small pores.
What is single digest vs Double Digest?
Definition. Single-digested plasmid refers to a plasmid digested by a single restriction enzyme while double-digested plasmid refers to a plasmid digested by two different restriction enzymes.
What is Thermo Scientific FastDigest SFII restriction enzyme?
Thermo Scientific FastDigest SfiI restriction enzyme recognizes GGCCNNNN^NGGCC site and cuts best at 50°C in 5–15 minutes using universal FastDigest Buffer. Thermo Scientific FastDigest SfiI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.
What is FastDigest SFII?
Thermo Scientific FastDigest SfiI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.
What is SFII E coli?
An E. coli strain that carries the SfiI gene from Streptomyces fimbriatus (ATCC 15051). One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 50°C in a total reaction volume of 50 µl. SfiI requires two copies of its recognition sequence for cleavage to occur.
What is the best restriction enzyme for DNA digestion?
Note: Also available as a FastDigest enzyme for rapid DNA digestion. Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions.