133,00 € – 538,00 €
TRANSCRIPTME is a modified, recombinant form of the Reverse Transcriptase from Moloney Murine Leukemia Virus (M-MuLV) purified from Escherichia coli. TRANSCRIPTME Reverse Transcriptase synthesizes a complementary DNA strand in the presence of a primer using either RNA (cDNA synthesis) or single-stranded DNA (ssDNA) as a template.
TRANSCRIPTME is a modified, recombinant form of the Reverse Transcriptase from the Moloney Murine Leukemia Virus (M-MuLV) purified from Escherichia coli. TRANSCRIPTME synthesizes a complementary DNA strand in the presence of a primer using either RNA (cDNA synthesis) or single-stranded DNA (ssDNA) as a template.
TRANSCRIPTME Reverse Transcriptase has increased thermal stability, which allows the reaction to be carried out at a higher temperature (optimum activity at 50°C). It increases the efficiency and specificity of those transcribed RNA regions which are rich in GC pairs and/or contain secondary structures. The enzyme has no 3’ –› 5’ exonuclease or RNase H activity, which improves the synthesis of a full-length cDNA, even from long mRNA templates, using random priming. The enzyme gives high yields of first strand cDNA up to 10 kb long.
Features and advantages
- High yields of full-length cDNA synthesis (up to 10 kb long)
- RNA and DNA-dependent DNA polymerase activities
- Increased sensitivity in RT-qPCR and RT-PCR assays
- Starting material: 10 pg – 5 g of total RNA or 10 pg – 500 ng of mRNA
- Optimal reaction temperature: 50°C
- Increased thermostability
- No RNase H or 3’–› 5’ exonuclease activities
- Suitable for the amplification of difficult RNA templates
- Full-length cDNA template synthesis for RT-qPCR and two-step RT-PCR assays
- cDNA synthesis for molecular cloning
- cDNA library construction
- RNA analysis
10 mM Tris-HCl (pH 8.0), 80 mM NaCl, 0.2mM EDTA, 50% (v/v) glycerol
10x RT Reaction Buffer
500 Tris-HCl (pH 8.3), 750 mM KCl, 30 mM MgCl2, 100 mM DTT
TRANSCRIPTME Reverse Transcriptase (RT32) - Manual
TRANSCRIPTME Reverse Transcriptase (RT32) – Troubleshooting
- We recommend the EXTRACTME TOTAL RNA KIT (EM09.1) and EXTRACTME TOTAL RNA PLUS KIT (EM11.1) for total RNA isolation from tissues and cell cultures
- During RT-PCR reaction mixture preparation, keep the TRANSCRIPTME Reverse Transcriptase and 10x RT Reaction Buffer on ice or in a freezing rack
- Use an RNase H treatment for reactions sensitive to residue RNA traces in order to increase the sensitivity of the RT-qPCR
- The quantity of cDNA used when preparing PCR or qPCR reactions should not exceed 1/10 of the final reaction volume; e.g. a maximum volume of 2.5 µl of cDNA should be used in a 25 µl reaction
- The activity of TRANSCRIPTME Reverse Transcriptaseis inhibited by metal ion chelating agents (e.g. EDTA), inorganic phosphor, pyrophosphate and polyamines
- Enzyme inactivation should be carried out at 85°C for 5 min.
Working with RNA
Acquisition of high quality, intact RNA, free of genomic DNA and RNase traces, is vital for the synthesis of a full-length cDNA followed by an accurate quantitative analysis (qPCR). The following recommendations for working with RNA should therefore be followed:
- Maintain aseptic working conditions: use disposable gloves, changing them as frequently, as required; use RNase-free consumables; only work in an area assigned for working with RNA and with equipment designated for that purpose.
- DNase I enzyme (not included) may be used if obtaining a DNA-free RNA sample is required.
- RNA samples should be stored aliquoted at -70°C. Avoid subjecting the samples to repeated freezing and thawing cycles.
The absence of DNase and RNase activities has been confirmed using the relevant procedures.
TRANSCRIPTME Reverse Transcriptase is > 90% pure as judged by SDS polyacrylamide gel. In addition, the functional quality is tested by RT-PCR experiment.
TRANSCRIPTME Reverse Transcriptase (RT32) - MSDS
TRANSCRIPTME Reverse Transcriptase (RT32) - CoA 1.2018 L.766772
TRANSCRIPTME Reverse Transcriptase (RT32) - CoA 1.2018 L.775873
TRANSCRIPTME Reverse Transcriptase (RT32) - CoA 2.2017 L.645672
TRANSCRIPTME Reverse Transcriptase (RT32) - CoA 1.2018 L.745673
Expression of Toll‐like receptors and costimulatory molecules in splenic B cells in a normal and abortion‐prone murine pregnancy model
Authors Daria Lorek, Anna Ewa Kedzierska, Anna Slawek, Anna Chelmonska‐Soyta Products TRANSCRIPTME Reverse Transcriptase (RT32) Year 2019 Source Am J Reprod Immunol. 2019 Aug;82(2)
A comprehensive transcriptome analysis of skeletal muscles in two Polish pig breeds differing in fat and meat quality traits
Authors Katarzyna Piórkowska, Kacper Żukowski, Katarzyna Ropka-Molik, Mirosław Tyra and Artur Gurgul Products TRANSCRIPTME Reverse Transcriptase (RT32) Year 2018 Source Genetics and Molecular Biology
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 (RT32) Year 2017 Source Histochem Cell Biol (2017) 148:105–115
Oligonucleotide gap-ﬁll ligation for mutation detection and sequencing in situ
Authors Marco Mignardi, Anja Mezger, Xiaoyan Qian, Linnea La Fleur, Johan Botling, Chatarina Larsson and Mats Nilsson Products TRANSCRIPTME Reverse Transcriptase (RT32) Year 2015 Source Nucleic Acids Research, 2015, Vol. 43, No. 22