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TRANSRTRO

Roche

Transcriptor Reverse Transcriptase

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Synonym(s):
RT-PCR, reverse transcriptase

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)

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This Item
TOSRTROTHIFICDNARO11855476001
vibrant-m

TOSRTRO

Transcriptor One-Step RT-PCR Kit

vibrant-m

11855476001

Titan One Tube RT-PCR System

specific activity

0.05 U/mg

specific activity

-

specific activity

-

specific activity

-

assay

≥90% (SDS-PAGE)

assay

-

assay

-

assay

-

recombinant

expressed in E. coli

recombinant

-

recombinant

-

recombinant

-

suitability

suitable for molecular biology

suitability

-

suitability

-

suitability

-

application(s)

genomic analysis
life science and biopharma

application(s)

-

application(s)

-

application(s)

-

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.
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.

Components

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

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.

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.

Storage and Stability

Avoid repeated freezing and thawing.

Other Notes

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

Legal Information

LightCycler is a registered trademark of Roche

Storage Class

12 - Non Combustible Liquids

wgk_germany

WGK 1

flash_point_f

does not flash

flash_point_c

does not flash


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Related Content

RT-qPCR, or quantitative reverse transcription PCR, combines the effects of reverse transcription and quantitative PCR or real-time PCR to amplify and detect specific targets. RT-qPCR has a variety of applications including quantifying gene expression levels, validating RNA interference (RNAi), and detecting pathogens such as viruses.

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