What type of restriction enzyme is XmaI?
XmaI is an isoschizomer of SmaI.
What enzyme digests the fastest?
DNA/RNA modifying enzymes, such as ligases, phosphatases, kinases and mesophilic DNA polymerases have 100% activity in FastDigest and FastDigest Green Buffer.
What is double digestion?
Digesting a DNA substrate with two restriction endonucleases simultaneously (double digestion) is a common timesaving procedure. Selecting the best NEBuffer to provide reaction conditions that optimize enzyme activity as well as avoid star activity associated with some enzymes is an important consideration.
What is XmaI?
XmaI is an isoschizomer of SmaI. XmaI produces a 5´ extension whereas SmaI produces blunt-ended fragments. Impaired by CpG methylation. star activity may result from a glycerol concentration of >5%
What are type II restriction enzymes?
Type II restriction enzymes are the familiar ones used for everyday molecular biology applications such as gene cloning and DNA fragmentation and analysis. These enzymes cleave DNA at fixed positions with respect to their recognition sequence, creating reproducible fragments and distinct gel electrophoresis patterns.
How do you test for double digestion?
Digestion: 1ug of DNA+2uL of Cut smart buffer (10x if your using NEB product)+1uL of each enzyme (according to your mentioned stock conc.)+ rest make up with sterile milliQ. Incubate at 37oC for 3 hrs, estimate the digested products concentration. Verify that both enzymes using the same reaction buffer.
What are isoschizomers and Neoschizomers describe with relevant examples?
Isoschizomers are pairs of restriction enzymes specific to the same recognition sequence. For example, SphI (CGTAC/G) and BbuI (CGTAC/G) are isoschizomers of each other. Neoschizomers are a specific type (subset) of isoschizomer. For example, SmaI (CCC/GGG) and XmaI (C/CCGGG) are neoschizomers of each other.
Does Xma1 produce sticky ends?
the C (of the Xma1 substrate) are part of the substrate recognition sequence of, but they do not form part of the stick ends. The sticky ends formed by these two enzymes are actually identical.
When did Fermentas start making FastDigest restriction enzymes?
In 2006, the innovative Lithuanian biotech company Fermentas launched a new product line for molecular biology researchers: the FastDigest restriction enzymes. Together with the polymerase chain reaction (PCR), restriction enzymes (REs) are arguably the most important tools in molecular biology.
What are the isoschizomers for FastDigest SMAI?
Isoschizomers: TspMI, XmaCI, XmaI. Thermo Scientific FastDigest SmaI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.
How does Thermo Scientific FastDigest SMAI work?
Thermo Scientific FastDigest SmaI is one of an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers. The universal buffer allows rapid single-, double-, or multiple DNA digestion within 5–15 minutes eliminating any need for buffer change or subsequent DNA…
How are restriction enzymes used in MBI Fermentas?
FastDigest® Restriction Enzymes by MBI Fermentas. FastDigest® restriction enzymes are an advanced line of restriction enzymes for rapid DNA digestion. All FastDigest® enzymes are 100% active in the universal FastDigest® Green buffer and are able to digest DNA in 5-15 min.
