RNase T1
Detailní popis
Thermo Scientific RNase T1 is an endoribonuclease that specifically degrades single-stranded RNA at G residues (see Figure 1 in Supporting Data). It cleaves the phosphodiester bond between the 3'-guanylic residue and the 5'-OH residue of adjacent nucleotides with the formation of corresponding intermediate 2', 3'-cyclic phosphate (see Reference 1). The reaction products are 3'-GMP and oligonucleotides with a terminal 3'-GMP.
RNase T1 does not require metal ions for activity.
Applications
- Removal of RNA from DNA preparations
- RNA sequencing (see Reference 1)
- Ribonuclease protection assays. Used in conjunction with RNase A (see Reference 2)
- Removal of RNA from recombinant protein preparations
- Determination of the level of RNA transcripts synthesized in vitro from DNA templates containing a "G-less cassette" (see Reference 3)
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Concentration | 1000 U/µL |
Definition of Activity Unit | 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. |
Hazardous | No |
Hazardous: | No |
Inactivation | Inactivation by heating is reversible, reliably removed by spin column or phenol/chloroform extraction. |
Inhibition | Inhibitors: 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). |
Molecular Weight | 11.2 kDa monomer. |
Quality Control |
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Source | E. coli cells with a cloned rntA gene of Aspergillus oryzae. |
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Storage Buffer | The enzyme is supplied in 50 mM Tris-HCl (pH 7.4) and 50% (v/v) glycerol. |
Storage Condition | -20 C |
Storage Condition: | -20 C |
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