RNase Inhibitor（RNasin）

Cat. No.: R292575

GRADE & PURITY 40U/μl

Descripción general

This product is a recombinant protein expressed in Escherichia coli and forms a 1:1 complex with RNase A. It exhibits a high degree of non-competitive inhibition of RNase. This product is of protein nature and is compatible with other competitive inhibitors (nucleic acids, inorganic phosphates). ) Different, it can be easily removed from the reaction system by phenol treatment.

Product content

RNase Inhibitor (RNasin) (40U/μl)

Purity

1) 300 U of RNasin and 1 μg of λDNA-Hind III decomposition product were reacted at 37°C for 1 hour, and the DNA electrophoresis band did not change.

2) 300U RNasin and 1μg Closed circμlar (RFI) pBR322 DNA are reacted at 37°C for 1 hour, and the DNA electrophoresis band does not change.

3) 100 U of RNasin and 1 μg of 16S, 23S rRNA are reacted at 37°C for 1 hour, and the electrophoresis band of RNA does not change.

5) SDS-PAGE: a single band at a molecular weight of 64KDa.

Application

1) cDNA synthesis reaction;

2) Polysome isolation;

3) In vitro translation;

4) In vitro transcription of SP6 or T7 RNA polymerase

5) In vitro transcription with cell-free extract.

Use attention

1) The pH range of inhibitory activity is wide, and the maximum activity is shown at pH 7～8;

2) Foaming or strong stirring (Vortex, etc.) will cause inactivation;

3) Does not inhibit RNase H activity.

Use case

Synthesis of the first strand of cDNA.

1) Prepare the following reaction solution in a microcentrifuge tube, the total volume is 20μl

Reagent	Volume	Final concentration
Total RNA or Poly（A）RNA	1-5μg	—
5×MMLV RT Buffer	4μl	1×
dNTP Mixture（25mM ）	0.4μl	0.5mM
Oligo dT or Random Primer （20μM）	1μl	1μM
RNasin（40U/μl）	1μl	2U/μl
MMLV Reverse Transcriptase（200U/μl）	1μl	—
Ultra-pure water	Up to 20μl	—
Total capacity	20μl	—

2) Stir gently.

3) After placing it at room temperature for 10 minutes, move it into a 42°C temperature bath.

4) Incubate at 42°C for 1 hour.

5) Move the sample into ice and cool for 2 minutes.

Specifications

Product Name

RNase Inhibitor（RNasin）

Especificaciones y pureza

40U/μl

Tipo de molécula

Small molecule

Almacenamiento y envío

Concentración

40U/μl

Condiciones de almacenamiento de almacenamiento

Store at -20°C

Enviado en

Ice chest + Ice pads

Definición de unidad

The amount of enzyme required to inhibit 50% of the activity of 5 ng RNase A is defined as 1 activity unit (U). The activity of RNase A is determined quantitatively by hydrolyzing Cyclic2′, 3′-CMP to generate 3′-CMP.