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11119915001 Roche

RNase, DNase-free

from bovine pancreas

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Properties

Related Categories Application Index, Biochemicals and Reagents, Enzymes, Inhibitors, and Substrates, Molecular Biology Tested, Protein and Nucleic Acid Isolation More...
form   solution
specific activity activity   ≥30 units/mg protein
packaging   pkg of 500 μg (1 ml)
mfr. no.   Roche
shipped in   dry ice
storage temp.   −20°C

Description

General description

Pyrimidine-specific endoribonuclease that acts on single-stranded RNA. 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. Before use, most RNase preparations must be boiled to remove DNase activity. This preparation of RNase does not need to be boiled; it can be used directly from the vial.

Other Notes

For life science research only. Not for use in diagnostic procedures.

Preparation Note

Working concentration: The optimal working concentration for RNase, DNase free, is 2 to 5 μg/ml. The reaction volume will vary for different applications. Some suggested guidelines are given below:

  1. For small-scale isolation of plasmid DNA ("miniprep" from a 1.5 ml bacterial culture), use 0.5 μl of RNase, DNase-free in a reaction volume of 50 μl.
    • To isolate plasmid DNA from a 100 ml bacterial culture, use 8 μl of RNase, DNase-free in a reaction volume of 2 ml.
    • To isolate genomic DNA from cultured mammalian cells (5 x 107 cells), use 8 μl of RNase, DNase-free in a reaction volume of 2 ml.

Working solution: Storage and Dilution Buffer: 10 mM Tris-HCl, 5 mM CaCl2, 50% glycerol (v/v), pH 7.0.

Unit Definition

One Kunitz unit is the amount of enzyme that causes a decrease in absorbance of A0 to A1 within one minute under the assay conditions. A0 to A1 corresponds to the total conversion, A1 being the final absorbance.
One unit produces a decrease in absorbance at 260 nm, which is equivalent to a total conversion of RNA to oligonucleotides in one minute at +25 °C.

Physical form

Solution, 500 μg/ml, in 10 mM Tris-HCl, 5 mM CaCl2, 50% glycerol (pH 7.0).

Application

RNase, DNase-free, efficiently removes contaminating RNA from plasmid or genomic DNA preparations.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Documents

Certificate of Analysis

Protocols & Articles

Protocols

Analyzing real-time PCR data by the comparative CT method

Two different methods of presenting quantitative gene expression exist: absolute and relative quantification. Absolute quantification calculates the copy number of the gene usually by relating the PC...
Keywords: Amplification, Anti-inflammatory agents, Apoptosis, Cancer, Clinical, Dehydration reaction, Diagnostic, Gene expression, Hormones, Methylations, Microarray Analysis, Molecular biology, Polymerase chain reaction, Polymerase chain reaction - quantitative, Reductions, Transcription, Transduction, Transfection, cDNA microarrays, transformation

RNase, DNase-free Protocol

Conditions for RNase digestion 0.1 mU RNase, DNase-free degrades 1 μg RNA in 30 min at + 37 °C in a reaction volume of 50 μl PCR grade water. The protein concentration of RNase, DNase-free is 0.5 μg/...
Keywords: Digestions, Polymerase chain reaction

Peer-Reviewed Papers
15

References

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