Quality level: 100
- enough for 25 reactions
- enough for 50 reactions
- enough for 500 reactions
dNTP included: no
hot start: no
- 200 reaction pack (03531287001)
- 25 reaction pack (03531317001)
- 50 reaction pack (03531295001)
Detection method: probe-based
In retroviruses such as human immunodeficiency virus type 1 (HIV-1), reverse transcriptase (RT) is the main enzyme. HIV-1 RT is made up of two subunits of 66 kDa and 51 kDa (p66 and p5l). A human endogenous retrovirus of the HERV-K family encodes the enzyme reverse transcriptase (RT). A recombinant reverse transcriptase for robust transcription of RNA fragments up to 14 kb was used in two-step RT-PCR in thermal cyclers and real-time PCR instruments.
Transcriber 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 (eg, 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 labelling during cDNA synthesis
- Recovery and cloning of the 5 ′ and 3 ′ ends of mRNA by RACE
- Generation of cDNA libraries with large inserts.
- Dideoxy DNA sequencing
- RNA sequencing
- Marking the 3 ‘end of DNA fragments
- Generation of single-stranded probes for genomic fingerprints
- In reverse transcription of human papillomavirus E6 RNA, cortical and striatal tissues, muscle biopsies from
- Becker muscular dystrophy specimens, and human brain microvascular endothelial cell and oocyte miRNA stem-loop (HCMC)
Biochemical / Physiological Actions
The reverse transcriptase of human immunodeficiency virus type 1 (HIV-1 RT) is essential for the catalytic conversion of single-stranded viral RNA into linear double-stranded DNA that is integrated into host cell chromosomes.
Features and Benefits
- Achieve high sensitivity in two-step RT-PCR.
Reverse Transcriptase Transcriptor is used in conventional thermal cyclers and real-time PCR instruments (eg LightCycler® instruments).
- Get more complete transcripts, up to 14 kb.
Large insert cDNA libraries can be generated.
- Transcribe difficult templates in reverse.
The enzyme works well at elevated temperatures, thereby overcoming the secondary structure of RNA (eg, GC-rich RNA templates) and providing optimal reaction conditions.
- Efficiently label the cDNA.
Nucleotides labelled with Cy3, Cy5, DIG, biotin, or aminoallyl are incorporated during cDNA synthesis.
- Transcriber Reverse Transcriptase, in storage buffer
- Transcriptional Buffer RT, 5x concentrated
Each lot of Transcriptor Reverse Transcriptase is routinely tested in RT-PCR:
- with a conventional thermal cycler to detect a 10 kb fragment of the human dystrophin gene, from total RNA of human skeletal muscle
- with the LightCycler® instrument to detect 5×102 to 5×106 copies of human PBGD RNA transcribed in vitro. Results are defined at fixed crossover points and fixed fluorescence intensity.
Reverse transcriptase transcriber is recommended for RT-PCR of:
- long targets, because it can transcribe RNA templates up to 14 kb
- rare objectives, because it has a high sensitivity
- GC-rich lenses, because can operate at high temperatures (up to + 65oC) to eliminate the problems associated with a large secondary structure.
Labelling for many applications
Transcriber reverse transcriptase is also recommended for preparing labelled cDNA, as it accepts a wide variety of modified nucleotides (including Cy3, Cy5, DIG, biotin, or aminoallyl-labelled dNTPs).
Transcriptionist accepts ssRNA and ssDNA templates and requires a primer for transcription.
For life science research only. It should not be used in diagnostic procedures.
LightCycler is a registered trademark of Roche