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RNAi and RNA Enzymes >
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RNase If |
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Prices are in US dollars and valid only for US orders.
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- Isolated from a recombinant source
- Eliminates RNA from DNA and protein preparations
- Supplied with 10X Reaction Buffer
Description:
Ribonuclease If (RNase If) is a single strand specific RNA endonuclease which will cleave at all RNA dinucleotide bonds leaving a 5´ hydroxyl and 2´, 3´ cyclic monophosphate (1). RNase If is a recombinant protein fusion of RNase I (from E. coli) and maltose-binding protein. It has identical activity to RNase I.
Source:
An E. coli strain containing a genetic fusion of the RNase I gene (rna) from E. coli and the gene coding for maltose-binding protein (MBP)(2).
Applications:- Degradation of single-stranded RNA to mono-, di- and trinucleotide (3)
- Used in ribonuclease protection assays
Reagents Supplied:
NEBuffer 3 (10X)
Enzyme Properties
Heat Inactivation:
70°C for 20 minutes
Reaction & Storage Conditions
Reaction Conditions:
1X NEBuffer 3
Incubate at
37°C.
1X NEBuffer 3:
50 mM Tris-HCl
100 mM NaCl
10 mM MgCl2
1 mM Dithiothreitol
pH 7.9 @ 25°C
Unit Definition:
One unit is defined as the amount of enzyme required to fully digest 1 picomole of synthetic ssRNA 33-mer in a total reaction volume of 10 µl in 15 minutes in 1X NEBuffer 3 as visualized on a 20% acrylamine gel (40:1 Bis) stained with SYBR Gold®.
Concentration:
50,000 units/ml
Storage Conditions:
10 mM Tris-HCl
100 mM NaCl
1 mM Dithiothreitol
0.5 mM EDTA
50% Glycerol
pH 8.0 @ 25°C
Storage Temperature:
-20°C
Notes
General notes:- RNase If will not degrade DNA. It has a strong preference for single-stranded RNA over double-stranded RNA.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
Purified free of contaminating endonucleases and exonucleases.
SS DNA Exonuclease Activity:
Incubation of 50 units of enzyme with 1 μg sonicated and denatured
[3H]-DNA (105 cpm/μg) for 30 minutes at 25ºC
in 50 μl reaction buffer released < 1% radioactivity.
ds DNA Exonuclease Activity:
Incubation of 50 units of enzyme with 1 μg sonicated [3H] DNA (105 cpm/μg) for 30 minutes at 25°C in 50 μl reaction buffer released < 1% radioactivity.
Endonuclease Activity:
Incubation of 50 units of enzyme with 1 μg of ΦX174 RF I DNA for 1 hour at 25ºC in 50 μl reaction buffer resulted in < 10% conversion to RFII.
References
- Spahr, P.F. and Hollingworth, B.R. (1961) J. Biol. Chem., 236, 823-831.
- Meador, J. III and Kennell, D. (1990) Gene, 95, 1-7.
- Meador, J. III, Cannon, B., Cannistraro, V.J. and Kennell, D. (1990) Eur. J. Biochem., 187, 549-553.
Reagents Sold Separately
NEBuffer 3
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