RNase A/T1 Mix

Detailní popis

Thermo Scientific RNase A/T1 Mix combines the RNA degradation activity of both RNase A and RNase T1. The RNase A specifically hydrolyzes RNA at C and U residues; RNase T1 specifically hydrolyzes RNA at G residues (see Figure 1 in Supporting Data [see Reference 1]).

Highlights

• Higher level of RNA degradation than with RNase A and RNase T1 separately
• Free of DNase activity - It is not necessary to heat the mix before use

Applications

• Removal of RNA from DNA preparations (see Reference 2)
• Removal of RNA from recombinant protein preparations
• Ribonuclease protection assays (see​ Reference 2)

  Concentration	2 mg/mL (approx. 10,000 U/mL (200 Kunitz U/mL)) of RNase A; 5000 u/mL of RNase T1
  Hazardous	No
  Inactivation	Not inactivated by heating, reliably removed by spin column or phenol/chloroform extraction.
  Inhibition	Inhibitors for RNase A: the most potent inhibitor is a mamalian ribonuclease inhibitor, e.g., RiboLock RNase Inhibitor. Other inhibitors: uridine 2', 3'-cyclic vanadate, 5'-diphosphoadenosine 3'-phosphate, and 5'-diphosphoadenosine 2-phosphate (see Reference 2), SDS, diethyl pyrocarbonate, 4 M guanidinium thiocyanate plus 0.1 M 2-mercaptoethanol and heavy metal ions. Inhibitors for RNase T1: metal ions Mg2+, Ca2+, Zn2+, Fe2+, Cu2+ (MgCl2 at 100 mM concentration is approx. 40% inhibitory, CaCl2 at 10 mM is approx. 30% inhibitory); mononucleotides (2'-GMP, 3'-GMP, etc.); guanilyl-2',5'-guanosine is a specific inhibitor (see Reference 4).
  Quality Control	The absence of endo-, exodeoxyribonucleases, and proteases confirmed by appropriate quality tests. Functionally tested for degradation of RNA in the plasmid DNA isolation procedure.
  Source	RNase A: Bovine pancreas. RNase T1: E.coli cells with a cloned rntA gene of Aspergillus oryzae.
  Storage Buffer	The enzyme is supplied in 50 mM Tris-HCl (pH 7.4) and 50% (v/v) glycerol.
  Storage Condition	-20 C
  Unit Definition	RNase A: One unit of the enzyme causes an increase in absorbance of 1.0 at 260 nm when yeast RNA is hydrolyzed at 37°C and pH 5.0. Fifty units are approximately equivalent to 1 Kunitz unit (see Reference 3). RNase T1: One unit of the enzyme causes an increase in absorbance of 1.0 at 260 nm in 15 min when yeast RNA is hydrolyzed at 37°C and pH 7.5.