Appearance (Form)
Liquid
Appearance (Colour)
Colourless
Appearance (Clarity)
Clear
DNase activity
None detected
Protease activity
None detected
Concentration
5U/µl
Description
RNase H is a 18.9 kDa recombinant endoribonuclease purified from an Escherichia coli strain, which over-expresses cloned RNase H gene (rnh). The enzyme hydrolyses specifically the phosphodiester bonds of RNA hybridized to DNA and produces 5´ phosphate-terminated oligoribonucleotides and single-stranded DNA.
Rnase H does not degrade single and double-stranded DNA or unhybridized RNA. It is a key enzyme in the removal of mRNA after first-strand cDNA synthesis. Treating cDNA with RNase H prior to PCR can improve sensitivity as RNA bonded to the cDNA template may prevent binding of the amplification primers in a PCR reaction. RNase H treatment is often necessary when amplifying longer, full-length cDNA targets. In addition, RNase H is useful for the removal of poly(A) tails on mRNAs after hybridization with oligo(dT) and also for the site-specific enzymatic cleavage of RNA.
Applications
Removal of RNA after first strand cDNA synthesis (RT-PCR and qRT-PCR)
Removal of mRNA prior to synthesis of second strand cDNA
Removal of the poly(A) sequences of mRNA after hybridization with oligo(dT)
Site-specific cleavage of RNA
Studies of in vitro polyadenylation reaction products
Includes
50U
RNAse H (5U/mcl) - 10mcl
10 X RNase H Reaction Buffer - 40 mcl
250U
RNAse H (5U/mcl) - 50mcl
10 X RNase H Reaction Buffer - 200 mcl
