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TRANSRTRO

Roche

Transcriptor Reverse Transcriptase

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About This Item

Codice UNSPSC:
41106313
NACRES:
NA.21

Ricombinante

expressed in E. coli

Livello qualitativo

Saggio

≥90% (SDS-PAGE)

Forma fisica

liquid

impiego

sufficient for 25 reactions
sufficient for 50 reactions
sufficient for 500 reactions

Attività specifica

0.05 U/mg

Caratteristiche

dNTPs included: no
hotstart: no

Produttore/marchio commerciale

Roche

Confezionamento

pkg of 200 reactions (03531287001)
pkg of 25 reactions (03531317001)
pkg of 50 reactions (03531295001)

tecniche

RT-PCR: suitable
RT-qPCR: suitable

Colore

colorless

pH

7.2

Solubilità

water: miscible

Compatibilità

suitable for molecular biology

N° accesso UniProt

applicazioni

genomic analysis
life science and biopharma

Metodo di rivelazione

probe-based

Attività estranea

DNase activity, none detected
Nicking activity, none detected
RNase activity, none detected

Temperatura di conservazione

−20°C (−15°C to −25°C)

Descrizione generale

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.

Applicazioni

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)

Azioni biochim/fisiol

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.

Caratteristiche e vantaggi

  • 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.

Componenti

  • Transcriptor Reverse Transcriptase, in storage buffer
  • Transcriptor RT buffer, 5x concentrated

Qualità

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.

Nota sulla preparazione

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.

Stoccaggio e stabilità

Avoid repeated freezing and thawing.

Altre note

For life science research only. Not for use in diagnostic procedures.

Note legali

LightCycler is a registered trademark of Roche

Codice della classe di stoccaggio

12 - Non Combustible Liquids

Classe di pericolosità dell'acqua (WGK)

WGK 1

Punto d’infiammabilità (°F)

does not flash

Punto d’infiammabilità (°C)

does not flash


Certificati d'analisi (COA)

Cerca il Certificati d'analisi (COA) digitando il numero di lotto/batch corrispondente. I numeri di lotto o di batch sono stampati sull'etichetta dei prodotti dopo la parola ‘Lotto’ o ‘Batch’.

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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
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)
Identification of an active reverse transcriptase enzyme encoded by a human endogenous HERV-K retrovirus.
Berkhout B, et al.
Journal of Virology, 73(3), 2365-2375 (1999)
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)
Fernando E Padovan-Neto et al.
Nitric oxide : biology and chemistry, 83, 40-50 (2018-12-12)
In Huntington's disease (HD), corticostriatal and striatopallidal projection neurons preferentially degenerate as a result of mutant huntingtin expression. Pathological deficits in nitric oxide (NO) signaling have also been reported in corticostriatal circuits in HD, however, the impact of age and

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