What type of nuclease is Micrococcal nuclease?
This micrococcal nuclease is a stable liquid form of the enzyme derived from Staphylococcus aureus. Micrococcal nuclease (MNase) exhibits exo- and endo-5′-phosphodiesterase activities against DNA and RNA. This enzyme digests double-stranded, single-stranded, circular and linear nucleic acids.
What is the function of Micrococcal nuclease?
Micrococcal Nuclease is an endonuclease that preferentially digests single-stranded DNA or RNA, especially at AT- or AU-rich regions. The enzyme will also digest double-stranded DNA or RNA, making it an essential component of chromatin immunoprecipitation (ChIP) assays.
What is staphylococcal nuclease?
Staphylococcal (or micrococcal) nuclease or thermonuclease (SNase or Nuc) is a naturally-secreted nucleic acid degrading enzyme that participates in Staphylococcus aureus spread in the infected host. Purified Nuc protein can be used as an exogenous reagent to clear cellular extracts and improve protein purification.
What does S1 nuclease do?
S1 Nuclease is a single-strand-specific endonuclease that hydrolyzes single-stranded RNA or DNA into 5´ mononucleotides. The enzyme will hydrolyze single-stranded regions in duplex DNA such as loops and gaps. S1 Nuclease is stable at 65°C.
How does MNase seq work?
Then, MNase-seq uses the endo-exonuclease micrococcal nuclease to bind and cleave protein-unbound regions of DNA of eukaryotic chromatin, first cleaving and resecting one strand, then cleaving the antiparallel strand as well. The chromatin can be optionally crosslinked with formaldehyde.
What can micrococcal nuclease be used for?
Micrococcal Nuclease is ideal for: Preparation of double-stranded DNA fragments with 5′-OH and 3′-phosphate. Studies of chromatin structure. Degradation of nucleic acids in crude cell-free extracts.
What is the optimal pH for micrococcal nuclease?
It is purified from a recombinant E. coli strain that digests double-stranded, single-stranded, circular and linear nucleic acids. The enzyme is active in the pH range of 7.0 – 10.0, with optimal activity at pH 9.2 for both RNA and DNA substrates.
How are histones protected in a micrococcal nuclease?
Once the nucleosome is assembled, the DNA wrapped around the histones is protected from MNase digestion, while the linker arms are digested. This will yield a distinct protected band of 147 bp in length.
What is the amount of micrococcal nuclease used in lane 2?
Digestion of 1 µg of Lambda genomic DNA with Micrococcal Nuclease in a 3-fold dilution series. The amount of enzyme used in Lane 2 is defined as 1 gel unit. Lane M is the PCR Marker (NEB #N3234).