| T751065 | Component | 200 U | 1000 U | Storage | | T751065A | Thermostable RNase H | 200 U | 5×200 U | -20°C. Avoid freeze/thaw cycle | | T751065B | 10× RNase H Reaction Buffer | 1 mL | 5×1 mL | -20°C. Avoid freeze/thaw cycle |
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Thermostable RNase H (Thermostable RNase H) is an endoribonuclease that remains active at higher temperatures (above 65°C). Thermostable RNase H selectively recognizes and enzymatically cuts RNA strands in a heterozygous double strand of DNA, while remaining intact in the heterozygous double strand. ThermostableRNase H does not degrade single - or double-stranded RNA or DNA.
Product application
1. Preparation of high-precision RNA structure map and analysis of mRNA capping rate.
2. mRNA was removed from second strand synthesis of CDNA; Improve RT-qPCR efficiency.
3. Remove the Poly(A) end of mRNA in the presence of Oligo(dT).
4. Components in isothermal amplification experiment.
Activity definition
The amount of enzyme required to produce 1nmol ribonucleotides within 20 minutes in a 50 μl reaction system at 50°C using a 40 pmol fluorescently labeled 25 bp RNA:DNA hybrid as substrate was defined as 1 unit of activity (U).
Quality control
1. Detection of non-specific endonuclease activity
5 U of this enzyme was incubated with superhelix plasmid DNA at 37°C for 4 h, and no changes were detected by DNA electrophoresis.
2. DNase residue detection
The enzyme was incubated with double-stranded DNA substrate at 37° C for 16 h, and the double-stranded DNA substrate was detected by DNA electrophoresis.
3. RNase residue detection
After 5 U of this enzyme was incubated with 500 ng RNA at 37°C for 1h, more than 90% of the RNA remained intact by agarose gel electrophoresis.
4. Host DNA residue detection
The residual amount of Escherichia coli host cell DNA in this product is less than 10 copies /5 U.
Attention
1.Thermostable RNase H reaction buffer contains MgCl2. Target Rnas: When DNA hybrids and other single-stranded Rnas (total Rnas) are used with ThermostableRNase H at high temperatures, it is recommended to control the reaction time and temperature to reduce the degradation of single-stranded Rnas of metal ions.
2.Thermostable RNase H can be inactivated by adding protease K or excessive EDTA.
3. The best reaction temperature is >65° C, the highest is 95° C. Thermostable RNase H is three to four times more active at 65°C than at 37°C. 4. For your safety and health, please wear a lab coat and wear disposable gloves.
