How do you prepare RNase A?
To prepare a 10 mg/mL RNase A stock solution, dissolve 100 mg of RNase A in 10 mL of Tris-Cl (10 mM, pH 7.5)/NaCl (15 mM). Heat to 100ºC for 5 min and cool at room temperature. Store at −20ºC. For a working dilution of 2 μg/mL, mix 20 μL of RNase A stock solution with 100 mL of TNE for SISH.
How can I make something DNase-free?
RNase, an enzyme that breaks down RNA, and DNase, which breaks down DNA, are contaminants that can interfere with nucleotide research. DNase can be destroyed by autoclaving for 15 minutes at 121°C or by following any of the procedures listed below.
How do you boil RNase A?
Dissolve RNase A at a concentration of 10 mg/ml in 0.01 M sodium acetate (pH 5.2); heat the solution to 100oC for 15 minutes. Cool slowly to room temperature and adjust the pH by addition of 0.1 volume of 1 M Tris.Cl (pH 7.4).
What is DNase-free RNase?
RNase, DNase-free, is a heterogeneous mixture of ribonucleases that has been prepared free of deoxyribonuclease activity according to the current Quality Control procedures. RNase, DNase-free, is particularly well suited for use in DNA isolation procedures.
How do you prepare RNase free water?
Popular Answers (1)
- Get MilliQ (reverse osmosis purified) water.
- Add 1 ml DEPC (Diethylpyrocarbonate) per 1000 ml of MilliQ or double distilled water (i.e. to a final concentration 0.1%) and mix thoroughly.
- Let the DEPC-mixed water incubate for 12 hours at 37°C.
- Autoclave DEPC-mixed water for 15 minutes.
Why is RNase A boiled?
DNAses are much less resistant than RNAses you can get rid of any contamination by placing the vial with RNA in boiling water for 20 min.
What is DNase RNase and protease?
Thermo Scientific RNase A, DNase and protease-free is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. It cleaves the phosphodiester bond between the 5′-ribose of a nucleotide and the phosphate group attached to the 3′-ribose of an adjacent pyrimidine nucleotide.
Does RNase DNase free mean sterile?
The fact that we call our products not sterile even when tested for DNA DNA/RNA fragments, DNase, RNase absence is only related to the general definition of sterile (after the process of sterilization less that 1 on 1000000 micro organism may survive).
How does RNase A, DNase and protease free work?
Learn more Thermo Scientific RNase A, DNase and protease-free is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. It cleaves the phosphodiester bond between the 5′-ribose of a nucleotide and the phosphate group attached to the 3′-ribose of an adjacent pyrimidine nucleotide.
How is RNase A used to treat pancreatitis?
Treatment with RNase A is an optional step in the protocol to remove any residual RNA present. This high-quality enzyme preparation is purified from bovine pancreas and is DNase, protease and DNA-free. It is supplied as a 20 mg/ml solution in 50% glycerol, 50 mM Tris-Cl, pH 7.4.
When to use monarch RNase A for DNA purification?
Monarch RNase A degrades single-stranded RNA at C and U residues. This formulation is purified from cow pancreas, is free of contaminating protease or DNase activity, and contains no residual host DNA. It is used at 56°C in the Monarch Genomic DNA Purification Kit protocol but is active at any temperature between 20°C and 65°C.
Why is RNase A used in GdNA extraction?
Co-purification of RNA during gDNA extractions is a common problem that leads to the overestimation of DNA yield. The low alcohol binding conditions employed in the kit optimizes binding of gDNA alone, so RNA co-purification is extremely low; however, an RNase A treatment step is included in each protocol as an option for the user.