RNase H
Product Description:
Ribonuclease H (RNase H) from E. coli specifically degrades
only the RNA strand of an RNA:DNA hybrid, leaving the DNA strand and any
unhybridized RNA intact. RNase H does not degrade single-stranded or
double-stranded DNAs or RNAs and is not inhibited by ScriptGuard™ RNase Inhibitor, an RNase A-family inhibitor.
RNase H can be used for the following applications:
- Elimination of RNA prior to second-strand cDNA synthesis.
- Removal of poly(A) tails from messenger RNA hybridized to oligo(dT).
- Oligodeoxyribonucleotide-directed site-specific cleavage of RNA.
- Specific destruction of hybrid-arrested mRNAs for translation experiments.
- Diagnostic assays using NASBA® transcription-based amplification methods.
- Diagnostic assays based on the Cycling Probe Technology.
- Template-dependent probe technologies.
Product Performance:
Figure 1. RNase H hydrolyzes a long DNA:RNA hybrid.
One nanomole of poly(rA):poly(T) substrate was treated with varying
amounts of RNase H for 20 min at 37°C. Samples treated with 1 U of RNase
H (lane 2) and 0.5 U of RNase H (lane 3) show hydrolysis of the hybrid.
Figure 2. RNase H hydrolyzes a short DNA:RNA hybrid.
20 pmoles of a 22-b ssDNA oligo was hybridized to 10 pmoles of a
1.85-kb ssRNA at coordinates 883-904 and treated with 0.02 U of RNase H
for 1 hour at 37°C. RNase H treatment hydrolyzes the hybrid, releasing
two bands of ~940 and ~880 bases in length (lane 3).
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