Is EcoRI restriction endonuclease a protein?
EcoRI (pronounced “eco R one”) is a restriction endonuclease enzyme isolated from species E. coli. It is a restriction enzyme that cleaves DNA double helices into fragments at specific sites, and is also a part of the restriction modification system….
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Do restriction enzymes cut proteins?
A restriction enzyme is a protein that recognizes a specific, short nucleotide sequence and cuts the DNA only at that specific site, which is known as restriction site or target sequence. More than 400 restriction enzymes have been isolated from the bacteria that manufacture them.
What is the enzymic function of restriction enzymes?
Restriction enzyme, also called restriction endonuclease, a protein produced by bacteria that cleaves DNA at specific sites along the molecule. In the bacterial cell, restriction enzymes cleave foreign DNA, thus eliminating infecting organisms.
How do you choose restriction enzymes for cloning?
When selecting restriction enzymes, you want to choose enzymes that:
- Flank your insert, but do not cut within your insert.
- Are in the desired location in your recipient plasmid (usually in the Multiple Cloning Site (MCS)), but do not cut elsewhere on the plasmid.
How do you select restriction enzymes?
Do I need to heat inactivate restriction enzyme?
FAQ: Is it necessary to inactivate restriction enzymes after vector digestion? Inactivation of restriction endonucleases is generally not necessary, but in some cases it might increase the transformation efficiency.
What are the reaction conditions for Apai restriction enzymes?
Learn more Thermo Scientific ApaI restriction enzyme recognizes GGGCC^C sites and cuts best at 37°C in B buffer (isoschizomers: PspOMI). See Reaction Conditions for Restriction Enzymes for a table of enzyme activity, conditions for double digestion, and heat inactivation for this and other restriction enzymes.
How is Apai inhibited in Thermo Scientific reaction buffers?
To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations. Note: Incubation at 30°C results in a 2-fold increase in activity. ApaI is inhibited by salt concentrations above 50 mM.
When to use Thermo Fisher Scientific FastDigest Apai buffer?
Thermo Scientific FastDigest ApaI restriction enzyme recognizes GGGCC^C site and cuts best at 37°C in 5–15 minutes using universal FastDigest Buffer. Isoschizomers: PspOMI.
What is the half life of Apai in E coli?
An E. coli strain that carries the ApaI gene from Acetobacter pasteurianus sub. pasteurianus (ATCC 9432). One unit is defined as the amount of enzyme required to digest 1 µg of pXba DNA in 1 hour at 25°C in a total reaction volume of 50 µl. Incubation at 37°C results in 100% activity, however, the half-life of ApaI at 37°C is only 30 minutes.