Which enzymes have star activity?
Which enzymes have star activity?
This altered specificity has been termed “star activity”. It has been suggested that star activity is a general property of restriction endonucleases (1) and that any restriction endonuclease will cleave noncanonical sites under certain extreme conditions, some of which are listed below.
What is BsrGI?
Thermo Scientific Bsp1407I (BsrGI) restriction enzyme recognizes T^GTACA sites and cuts best at 37°C in Tango buffer (isoschizomers: BsrGI, BstAUI). All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions.
Does noti have star activity?
100% activity in rCutSmart Buffer. Time-Saver™ qualified for digestion in 5-15 minutes. Reduced star activity.
What are NEB enzymes?
With over 40 years of offering restriction enzymes to the research community, NEB has earned the reputation of being a leader in enzyme technologies. Working continuously to be worthy of that distinction, NEB strives to develop enzymes of the highest purity and unparalleled performance.
How can star activity be avoided?
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. Use only Mg2+ as the divalent cation in the reaction buffer.
Why is it called star activity?
It has been demonstrated that under extreme non-standard conditions, restriction endonucleases are capable of cleaving sequences which are similar but not identical to their defined recognition sequence. This altered or relaxed specificity has been termed “”star”” activity.
Is NotI a restriction enzyme?
Thermo Scientific NotI restriction enzyme recognizes GC^GGCCGC sites and cuts best at 37°C in O buffer (Isoschizomers: CciNI). 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 recognition sequence for NotI?
The type IIP restriction enzyme NotI from Nocardia otitidis-caviarum is a homodimer which recognizes the 8 basepair DNA sequence 5′- GC/GGCCGC -3′ and cleaves both strands of DNA to create 5′, 4-base cohesive overhangs (Qiang and Schildkraut, 1987).
What are high fidelity enzymes?
High Fidelity Restriction Enzymes have been engineered by exchanging functional amino acid residues and then screening for optimal mutants that perform under a wide range of conditions.
Which type of restriction enzyme can recognize the HIF binding sequence?
CCCGGG within the binding sequence is palindromic – only a restriction enzyme that recognizes a four and six base sequence can recognize this sequence within the HIF binding sequence.
What causes star activity?
Generally, star activity (i.e., the enzyme cuts a sequence that is an imperfect match to its known target) is caused by nonoptimal reaction conditions.(1),(2) These include: High concentration of glycerol (>5% v/v). High ratio of enzyme units to micrograms of DNA.
How do you know if your restriction digestion was successful?
If the digested product would be visible at a lower coordinate on the gel, it would have made things easy. You can amplify your digested fragment with primer beginning in the flankers region and with only 3-4 bp in the intern 8680 bp region. If you do not get PCR fradments, was the digestion successfully.
