What is RNase free DNase?
What is RNase free DNase?
DNase I (RNase-free) is a recombinant endonuclease that nonspecifically catalyzes the degradation of both single- and double-stranded DNA and DNA-RNA hybrids, producing 5′-phosphate and 3′-hydroxyl termini-containing oligonucleotides of varying lengths.
How do you give a DNase treatment?
Tip: As a rule of thumb for the DNase I digestion, use one unit of DNase I per 1 to 5 μg of total RNA in a 50 μl total volume incubated for 20 minutes at +25 to +37°C. After the additional DNase digestion step an additional purification of the RNA from the DNase I enzyme is mandatory.
How do you use DNase 1?
A Typical DNase I Reaction Protocol (M0303)
- Set up the following reaction on ice: COMPONENTS. 100 μl REACTION. RNA. ~ 10 μg RNA. DNase I Reaction Buffer (10X) 10 μl (1X) DNAse I (RNase-free)
- Incubate at 37°C for 10 minutes.
- Add 1 µl of 0.5 M EDTA (to a final concentration of 5 mM).
- Heat inactivate at 75°C for 10 minutes.
What is the difference between DNase and RNase?
In the laboratory, DNase I is required to remove DNA from samples used in mRNA expression assays, whereas RNase A is used to remove RNA from samples used for DNA analysis. DNase and RNase are important for modifying and metabolizing nucleic acid chains and can be used as disease markers [4–13].
How do you maintain an RNase-free lab?
Schedule For RNase Control
- Daily. Use Ambion’s certified RNase-free buffers and reagents.
- Weekly. Thoroughly clean lab benchtops, pipettors (see sidebar below) and tube racks with RNaseZap.
- Monthly.
- As Needed.
What is RNase and DNase?
The function of nucleases (DNases and RNases) includes the enzymatic breakdown of DNA and RNA and is necessary for numerous research applications. For example, the purification of proteins and specific nucleic acids often requires the digestion of DNA, RNA or both.
Does DNase and RNase free mean sterile?
The exact defnition of something being RNAse free and non-sterile, means that the absence of RNAse has been validated but after sterilization the SAL value of the product did not met the requirement of 1/1000000. You can use these tubes for RNA work, because they are RNAse free.
What is DNase used for?
In molecular biology, DNase (namely DNase I) is used to degrade DNA in applications such as RNA isolation, reverse transcription preparation, DNA-protein interactions, cell culture, and DNA fragmentation. Clinical uses of DNase include breaking down of mucus to clear up respiratory tracts.