Products

Product Introduction:
This product is a recombinant protein expressed in Escherichia coli. It forms a 1:1 complex with RNase A and exhibits highly non-competitive inhibition of RNases (Ki = 3×10⁻¹⁰ M). The reaction is reversible: the complex can be dissociated by urea and thiol-containing reagents, resulting in the reactivation of RNase and the irreversible inactivation of the inhibitor. As a proteinaceous inhibitor, it differs from other competitive inhibitors (nucleic acid-based, inorganic phosphate-based) and can be easily removed from the reaction system by phenol treatment.

Activity Definition:
1.One activity unit (U) is defined as the amount of enzyme required to inhibit 50% of the activity of 5 ng RNase A.
2.First-Strand cDNA Synthesis (Taking 20 μL Reaction System as Example)

1) Add the Following Components
Components Volume
Total RNA/mRNA 50 ng-5 μg/5-500 ng
Oligo (dT) 18 (0.5 μg /μL) or
Random Primer (0.1 μg/μL) or
GSP (Gene Specific Primer) 1 μL
1 μL
2 pmol
dNTP Mixture (10 mM each) 1 μL
5× RT Buffer 4 μL
RNase Inhibitor (40 units/μL) 0.5-1 μL
TUREscript Hˉ RTase 1 μL
RNase free H2O to final volume 20 μL
2) Incubation Conditions
3) Mix gently.
If using Oligo (dT)₁₈ or Gene-Specific Primer (GSP), incubate at 42°C for 50 minutes.
If using Random Primer, incubate at 25°C for 10 minutes, then at 42°C for 50 minutes.
Heat at 70°C for 5 minutes to inactivate TUREscript H⁻ RTase.

3.RT-PCR
The cDNA obtained by reverse transcription can be directly used for PCR reactions or stored at -20°C.