What are the different types of restriction enzymes?
Traditionally, four types of restriction enzymes are recognized, designated I, II, III, and IV, which differ primarily in structure, cleavage site, specificity, and cofactors.
What is star activity restriction enzymes?
Star activity is the relaxation or alteration of the specificity of restriction enzyme mediated cleavage of DNA that can occur under reaction conditions that differ significantly from those optimal for the enzyme.
What are isoschizomers and Neoschizomers describe with relevant examples?
Isoschizomers are the restriction enzymes which recognize and cleave at the same recognition site. For example, SphI (CGTAC/G) and BbuI (CGTAC/G) are isoschizomers of each other. Neoschizomers are the restriction enzymes which recognize the same site and have a different cleavage pattern.
What do you mean by isoschizomers?
Isoschizomers (same cut) are restriction endonucleases that recognize the same DNA sequence and make the same cut. From: Molecular Biology, 1995.
Does BamHI have star activity?
After 50-fold overdigestion with BamHI, >95 % of the DNA fragments can be ligated and recut with this enzyme. Star activity: Conditions of low ionic strength, high enzyme concentration, glycerol concentration >5 % or pH >8.0 may result in star activity.
What is the restriction site for PstI?
Thermo Scientific PstI restriction enzyme recognizes CTGCA^G sites and cuts best at 37°C in O buffer (Isoschizomers: BspMAI). 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.
What is the difference between isoschizomers and Neoschizomers?
Isoschizomers are restriction enzymes that have the same recognition sequence and cleave the DNA at the same positions, while neoschizomers are restriction enzymes that have the same recognition sequence but cleave DNA at different positions. So, this is the key difference between isoschizomers and neoschizomers.
What is the restriction site for BamHI?
BamHI (from Bacillus amyloli) is a type II restriction endonuclease, having the capacity for recognizing short sequences (6 b.p.) of DNA and specifically cleaving them at a target site. This exhibit focuses on the structure-function relations of BamHI as described by Newman, et al.
How do you inactivate BamHI?
Only small amounts of BamHI (up to 10 units) can be inactivated at 80°C in 20 min. To prepare the digested DNA for electrophoresis: – stop the digestion reaction by adding 0.5 M EDTA, pH 8.0 (#R1021), to achieve a 20 mM final concentration. Mix thoroughly, add an electrophoresis loading dye and load onto gel.
Are isoschizomers interchangeable?
Isoschizomers are interchangeable, and neoschizomers are not, because they cut differently even if they recognize the same sequence.