TRANSRTRO
Roche
Transcriptor Reverse Transcriptase
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About This Item
UNSPSC Code:
41106313
NACRES:
NA.21
recombinant
expressed in E. coli
Quality Level
Assay
≥90% (SDS-PAGE)
form
liquid
usage
sufficient for 25 reactions
sufficient for 50 reactions
sufficient for 500 reactions
specific activity
0.05 U/mg
feature
dNTPs included: no
hotstart: no
manufacturer/tradename
Roche
packaging
pkg of 200 reactions (03531287001)
pkg of 25 reactions (03531317001)
pkg of 50 reactions (03531295001)
technique(s)
RT-PCR: suitable
RT-qPCR: suitable
color
colorless
pH
7.2
solubility
water: miscible
suitability
suitable for molecular biology
UniProt accession no.
application(s)
genomic analysis
life science and biopharma
detection method
probe-based
foreign activity
DNase activity, none detected
Nicking activity, none detected
RNase activity, none detected
storage temp.
−20°C (−15°C to −25°C)
Related Categories
General description
A recombinant reverse transcriptase for the robust transcription of RNA fragments up to 14 kb used in two-step RT-PCR on thermal cyclers and real-time PCR instruments.
In retroviruses like the human immunodeficiency virus type 1 (HIV-1), reverse transcriptase (RT) is the core enzyme. HIV-1 RT is made of two subunits of 66 kDa and 51 kDa (p66 and p5l). A human endogenous retrovirus of the HERV-K family codes for reverse transcriptase (RT) enzyme.
Application
Transcriptor Reverse Transcriptase is designed to transcribe RNA (mRNA, total RNA, viral RNA, and in vitro-transcribed RNA) from a variety of sources, using conventional thermal cyclers and real-time PCR instruments (e.g., the LightCycler® Instruments) for the following applications:
- Synthesis of first-strand cDNA for use in subsequent amplification reactions
- RT-PCR of GC-rich RNA templates
- Cy3, Cy5, DIG, biotin, and aminoallyl labeling during cDNA synthesis
- Retrieving and cloning the 5′ and 3′ termini of mRNA by RACE
- Generation of cDNA libraries with large inserts
- Dideoxy DNA sequencing
- RNA sequencing
- 3′-end labeling of DNA fragments
- Generation of single-stranded probes for genomic footprints
- In the reverse transcription of RNA from human Papillomavirus E6, cortical and striatal tissues, muscle biopsies of Becker muscular dystrophy samples and miRNA stem-loop from oocytes and human cerebral microvascular endothelial cells (hCMVECs)
Biochem/physiol Actions
Human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT) is essential for the catalytic conversion of single-stranded viral RNA into the double-stranded linear DNA that is integrated into host cell chromosomes.
Features and Benefits
- Achieve high sensitivity in two-step RT-PCR.
- Obtain more full-length transcripts - up to 14 kb.
- Reverse transcribe difficult templates.
- Efficiently label cDNA.
Preparation Note
Volume activity: 20 U/μl.
Transcription of long, rare, or difficult RNA targets
Transcriptor Reverse Transcriptase is recommended for RT-PCR of:
Labeling for many applications
Transcriptor Reverse Transcriptase is also recommended for preparing labeled cDNA, since it accepts a wide variety of modified nucleotides (including Cy3-, Cy5-, DIG-, Biotin-, or aminoallyl-labeled dNTPs).
Reaction requirements
Transcriptor accepts ssRNA and ssDNA templates and requires a primer for transcription.
Transcription of long, rare, or difficult RNA targets
Transcriptor Reverse Transcriptase is recommended for RT-PCR of:
- long targets, because it can transcribe up to 14 kb RNA templates
- rare targets, because it has high sensitivity
- GC-rich targets, because it can operate at high temperatures (up to +65oC) to eliminate problems associated with extensive secondary structure
Labeling for many applications
Transcriptor Reverse Transcriptase is also recommended for preparing labeled cDNA, since it accepts a wide variety of modified nucleotides (including Cy3-, Cy5-, DIG-, Biotin-, or aminoallyl-labeled dNTPs).
Reaction requirements
Transcriptor accepts ssRNA and ssDNA templates and requires a primer for transcription.
Avoid repeated freezing and thawing.
Analysis Note
Each lot of Transcriptor Reverse Transcriptase is routinely function tested in RT-PCR:
- with a conventional thermal cycler to detect a 10-kb fragment from the human dystrophin gene, starting from human skeletal muscle total RNA
- with the LightCycler® Instrument to detect 5 x 102 to 5 x 106 copies of in vitro-transcribed human PBGD RNA. The results are defined in fixed crossing points and fixed fluorescence intensity.
Other Notes
- Transcriptor Reverse Transcriptase, in storage buffer
- Transcriptor RT buffer, 5x concentrated
For life science research only. Not for use in diagnostic procedures.
Legal Information
LightCycler is a registered trademark of Roche
Storage Class Code
12 - Non Combustible Liquids
WGK
WGK 1
Flash Point(F)
does not flash
Flash Point(C)
does not flash
Regulatory Information
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Pietro Spitali et al.
Journal of cachexia, sarcopenia and muscle, 9(4), 715-726 (2018-04-24)
Analysis of muscle biopsies allowed to characterize the pathophysiological changes of Duchenne and Becker muscular dystrophies (D/BMD) leading to the clinical phenotype. Muscle tissue is often investigated during interventional dose finding studies to show in situ proof of concept and
Expression profiles and function analysis of microRNAs in postovulatory aging mouse oocytes
Wang TY, et al.
Aging, 9, 1186-1186 (2017)
Tracking disease progression non-invasively in Duchenne and Becker muscular dystrophies
Spitali P, et al.
Journal of Cachexia, Sarcopenia and Muscle, 9, 715-726 (2018)
Age-and sex-related changes in cortical and striatal nitric oxide synthase in the Q175 mouse model of Huntington's disease
Padovan-Neto FE, et al.
Nitric Oxide, 83, 40-50 (2019)
Crystal structure of human immunodeficiency virus type 1 reverse transcriptase complexed with double-stranded DNA at 3.0 A resolution shows bent DNA.
Jacobo-Molina A, et al.
Proceedings of the National Academy of Sciences of the USA, 90(13), 6320-6324 (1993)
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