Transcriptor Reverse Transcriptase
ROCHE/TRANSRTRO
Product Type: Chemical
Figure 1: Detection of mouse β-actin mRNA by two-step RT-PCR on a conventional thermal cycler.
Figure 2: Sensitivity in one-step RT-PCR of a 2-kb dystrophin RNA fragment (comparison of Transcriptor to other reverse transcriptases), with Expand High Fidelity Enzyme Blend used as the PCR component. Transcriptor Reverse Transcriptase exhibited the highest sensitivity.
Figure 3: Overcome difficult RNA templates by using Transcriptor Reverse Transcriptase at up to +65 °C. Transcriptor Reverse Transcriptase or other reverse transcriptases were used with a specific primer to reverse transcribe a 4.9-kb fragment (GC content: 70%) of human 28S ribosomal RNA at +55 °C, +60 °C, and +65 °C.
Figure 4: Produce full-length transcripts up to 14 kb for sensitive, high-yielding RT-PCR with Transcriptor Reverse Transcriptase. See Figure 5 for details.
Figure 5: Produce full-length transcripts up to 14 kb for sensitive, high-yielding RT-PCR with Transcriptor Reverse Transcriptase.
| application(s) | genomic analysis life science and biopharma |
| assay | ≥90% (SDS-PAGE) |
| color | colorless |
| detection method | probe-based |
| feature | dNTPs included: no |
| hotstart: no | |
| foreign activity | DNase activity, none detected |
| Nicking activity, none detected | |
| RNase activity, none detected | |
| form | liquid |
| manufacturer/tradename | Roche |
| packaging | pkg of 200 reactions (03531287001) |
| pkg of 25 reactions (03531317001) | |
| pkg of 50 reactions (03531295001) | |
| pH | 7.2 |
| Quality Level | 100  |
| recombinant | expressed in E. coli |
| solubility | water: miscible |
| specific activity | 0.05 U/mg |
| storage temp. | −20°C (−15°C to −25°C) |
| suitability | suitable for molecular biology |
| technique(s) | RT-PCR: suitable |
| RT-qPCR: suitable | |
| UniProt accession no. | Q9JG39  |
| usage | sufficient for 25 reactions |
| sufficient for 50 reactions | |
| sufficient for 500 reactions |
| 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. |
| Components: | • Transcriptor Reverse Transcriptase, in storage buffer • Transcriptor RT buffer, 5x concentrated |
| Features and Benefits: | • Achieve high sensitivity in two-step RT-PCR. Transcriptor Reverse Transcriptase is used in conventional thermal cyclers and real-time PCR instruments (e.g., the LightCycler®Instruments). • Obtain more full-length transcripts - up to 14 kb. cDNA libraries with large inserts can be generated. • Reverse transcribe difficult templates. The enzyme works well at elevated temperatures, thereby overcoming RNA secondary structure (e.g., GC-rich RNA templates) and facilitating optimal reaction conditions. • Efficiently label cDNA. Cy3-, Cy5-, DIG-, biotin-, or aminoallyl-labeled nucleotides are incorporated during cDNA synthesis. |
| 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. |
| General description: | 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. |
| Legal Information: | LightCycler is a registered trademark of Roche |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Preparation Note: | Volume activity: 20 U/μl. 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. |
| Quality: | 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. |
| Storage and Stability: | Avoid repeated freezing and thawing. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| Purity | ≥90% (SDS-PAGE) |
| activity | specific activity: 0.05 U/mg |
| Storage Temp. | −20°C (−15°C to −25°C) |
| UNSPSC | 41106313 |