How do you make RNase A stock solution?

How do you make RNase A stock solution?

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.

How long does RNase A last?

We recommend that Buffer P1 with RNase A be stored in the refrigerator (2-8°C). RNase A will be stable for 6 months under this condition.

How do you use RNase solution?

You simply spray RNaseZap® Solution onto the surface to be decontaminated and rinse it off with RNase-free water. Working with RNA requires that special measures be taken to ensure an RNase-free environment.

How do I get rid of RNase?

Soak in a 0.1% Aqueous Solution of Diethyl Pyrocarbonate (DEPC) for 2 hours at 37°C. Then, rinse several times with Sterile (DEPC-treated) Water***, Heat to 100°C for 15 minutes OR Autoclave for 15 minutes at 121°C on a Liquid/Slow Exhaust Cycle. (Heating or autoclaving will remove DEPC residues.)

How do you remove RNase solution?

DEPC destroys enzymatic activity by modifying -NH, -SH, and -OH groups in RNases and other proteins. The treatment typically involves incubating the solution at room temperature with 0.1% DEPC for a few hours, usually overnight, followed by autoclaving the solution to eliminate residual DEPC.

Why is RNase used?

RNase A is an endoribonuclease that specifically hydrolyzes RNA 3´ of pyrimidine residues and cleaves the phosphodiester linkage to the adjacent nucleotide. RNase A is used to remove RNA during procedures for the isolation of plasmid and genomic DNA.

How much RNase A should I use?

Recommended concentration of RNase A is 1 to 100 µg/mL depending on the application. The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 to 100 mM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well the RNA strand in RNA-DNA hybrids.

How do you store RNase A?

Store at RNase A at –20 °C. Stock solutions stored in frozen aliquots remain active for at least 6 months.