What is a fast digest restriction 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.
What is EcoRI buffer?
Thermo Scientific 10X Buffer EcoRI is the optimal buffer recommended for use with EcoRI restriction enzyme and is premixed with BSA for enhanced stability.
Is the ecor1 Restriction enzyme?
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.
Does ecor1 have star activity?
It has long been known that as with many other type II restriction endonucleases EcoRI is capable of cleaving “star” sequences with low activity (Polisky et al., 1975; Goodman et al., 1977; Tikchonenko et al., 1978; Hsu and Berg, 1978; Malyguine et al., 1980; Gardner et al., 1982; Rosenberg and Greene, 1982; Lesser et …
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 does R stand for in EcoRI?
EcoRI is a restriction endonuclease that is isolated from the bacterium Escherichia coli. In EcoRI, Eco represents the species of bacteria from which it is isolated i.e. Escherichia coli. R represents the strain of the bacteria which is RY-13 in this case.
What is the likely constituents of ecor1 buffer?
Storage buffer: EcoRI is supplied in 300 mM NaCl, 5 mM KPO4 (pH 7.4), 0.1 mM EDTA, 1 mM dithiothreitol, 0.15% Triton, 200 μg/ml BSA and 50% glycerol. Inactivation: Heat to 65 °C for 20 minutes.
What is the function of EcoR1?
What is the use of EcoRI? EcoRI is a type II restriction enzyme, which cuts the DNA specifically at the restriction site forming sticky ends. It makes the ligation reaction more efficient.
How does the target sequence differ between EcoRI and HindIII?
Thus, this is the key difference between EcoRI and HindIII. Furthermore, EcoRI cuts the DNA at the specific recognition sequence G↓AATTC, while HindIII cuts the DNA at the specific recognition sequence A↓ACCTT.
How can we prevent star activity in restriction digestion?
To prevent star activity, we recommend the following guidelines: Use as few units of restriction enzyme as possible for a complete digestion, which avoids overdigestion of the DNA and reduces the final glycerol concentration in the reaction.