85,00 € – 336,00 €
RNase H is a 18.9 kDa recombinant endoribonuclease purified from an Escherichia coli strain. The enzyme hydrolyses specifically the phosphodiester bonds of RNA hybridized to DNA and produces 5´ phosphate-terminated oligoribonucleotides and single-stranded DNA. The enzymes 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 treatment is often necessary when amplifying longer, full-length cDNA targets. In addition, this enzymes 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.
- Recombinant endoribonuclease over-expressed in E. coli.
- Hydrolyses specifically the phosphodiester bonds of RNA hybridized to DNA.
- Does not degrade single and double-stranded DNA or un-hybridized RNA.
- 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.
- 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.
One unit catalyses the hydrolysis of 1 nmol of RNA in [3H]-labeled poly(A)×poly(dT) to acid-soluble ribonucleotides in a total reaction volume of 50 µl in 20 min at 37°C in 1x Reaction Buffer.
Protocol & Manual
RNase H (RT34) - Manual
Use 5 U of enzyme to remove RNA from a RNA:DNA duplex after reverse transcription in a 20 μl reaction. If 50 μl reaction is desired, the use of 12.5 U of enzyme is recommended.
The reaction mixture should be incubated at 37⁰C for 20 minutes.
Store at -80°C or -20°C.
Shipping on dry or blue ice.
20 mM Tris-HCl (pH 7.9), 100 mM KCl, 10 mM MgCl2, 0.1 mM EDTA, 0.1 mM DTT, 50% (v/v) glycerol.
10x Reaction Buffer
200 mM Tris-HCl (pH 8.4), 500 mM KCl, 50 mM MgCl2, 200 mM DTT
- The activity of RNase H is inhibited by metal chelators (e.g. EDTA) and sulfhydryl SH-blocking reagents.
- Inactivate enzyme by heating at 65°C for 10 min.
Quality & Safety
RNase H is >90% pure as judged by SDS polyacrylamide gel. The absence of DNase activity has been confirmed using the relevant procedures. In addition, the functional quality is tested by RT and subsequent PCR and qPCR assays.
RNase H (RT34) - MSDS
RNase H (RT34) - CoA 1.2018 L.745673
A novel mutation tolerant padlock probe design for multiplexed detection of hypervariable RNA viruses
Authors: Sibel Ciftci, Felix Neumann, Iván Hernández-Neuta, Mikhayil Hakhverdyan, Ádám Bálint, David Herthnek, Narayanan Madaboosi & Mats Nilsson Products: RNase H (RT34), dNTPs, T4 DNA Ligase (EN11), ATP Year: 2019 Source: Scientific Reports, 2019, Vol 9, Article number: 2872
Targeted DNA sequencing and in situ mutation analysis using mobile phone microscopy
Authors: Malte Kühnemund Qingshan Wei, Evangelia Darai, Yingjie Wang, Iván Hernández-Neuta, Zhao Yang, Derek Tseng, Annika Ahlford, Lucy Mathot, Tobias Sjöblom, Aydogan Ozcan & Mats Nilsson Products: RNase H (RT34), TRANSCRIPTME RNA Kit (RT31), T4 DNA Ligase (EN11), dNTPs Year: 2017 Source: NATURE COMMUNICATIONS
Analysis of IAV Replication and Co-infection Dynamics by a Versatile RNA Viral Genome Labeling Method
Authors: Dan Dou, Iván Hernández-Neuta, Hao Wang, Henrik Östbye, Xiaoyan Qian, Swantje Thiele, Patricia Resa-Infante, Nancy Mounogou Kouassi, Vicky Sender, Karina Hentrich, Peter Mellroth, Birgitta Henriques-Normark, Gülsah Gabriel, Mats Nilsson and Robert Daniels Products: RNase H (RT34), TRANSCRIPTME RNA Kit (RT31) Year: 2017 Source: Cell Reports 20, 251–263, July 5, 2017
Visualization of tumor heterogeneity by in situ padlock probe technology in colorectal cancer
Authors: Amin El‑Heliebi, Karl Kashofer, Julia Fuchs, Stephan W. Jahn, Christian Viertler, Andrija Matak, Peter Sedlmayr, Gerald Hoefler Products: RNase H (RT34), TranscriptMe Reverse Transcriptase Year: 2017 Source: Histochem Cell Biol (2017) 148:105–115