10109118001

Reverse Transcriptase AMV

solution, >50 units/μg protein, suitable for RT-qPCR, suitable for RT-PCR

• Synonym(s): amv reverse transcriptase
• Enzyme Commission number: 2.7.7.49 (BRENDA, IUBMB)
• UNSPSC Code: 41105600

Properties

• form: solution
• Quality Level: 100
• specific activity: >50 units/μg protein
• feature: dNTPs included: no; hotstart: no
• packaging: pkg of 1,000 U
• manufacturer/tradename: Roche
• storage condition: avoid repeated freeze/thaw cycles
• parameter: 42 °C optimum reaction temp.
• technique(s): RT-PCR: suitable; RT-qPCR: suitable
• input: purified RNA
• detection method: probe-based

Description

General description

Reverse Transcriptase, AMV is a gene product of the RNA genome of avian myeloblastosis virus. The enzymatically active forms of the purified enzyme are α,ββ and αβ. The molecular weight of the α-subunit is 68 kDa, that of the β-subunit 92 kDa. The mature αβ form, the most active form of Reverse Transcriptase, AMV, includes a RNA-directed DNA polymerase, a DNA-dependent DNA polymerase, a RNase H, and an unwinding activity. Reverse Transcriptase, AMV is used for cDNA synthesis, for synthesis of first strand cDNA for use in subsequent amplification reactions and dideoxy DNA sequencing.
The enzyme can also be used for RNA sequencing, 3′ end labeling of DNA fragments, and the generation of ss probes for genomic footprints.

Reverse Transcriptase AMV requires a primer and Mg²⁺ or Mn²⁺ for activity.

Specificity

AMV (Avian Myeloblastosis Virus) Reverse Transcriptase is an RNA-directed DNA polymerase and a DNA-dependent DNA polymerase. The AMV reverse transcriptase requires a primer and Mg²⁺ or Mn²⁺ for activity.
Heat inactivation: 5 min, 95 °C

Application

Reverse Transcriptase AMV is suitable for:

• First- and second-strand cDNA synthesis and synthesis of first strand cDNA for use in subsequent amplification reactions (RT-PCR)
• Dideoxy DNA sequencing
• Primer extension
• RNA sequencing
• 3′-end labeling of DNA fragments
• Generation of single-stranded probes for genomic footprint experiments

Features and Benefits

• Efficiently transcribes total RNA, mRNA, viral RNA and RNA rich in secondary structures
• Procure full length cDNA fragments up to 12 kb
• Higher thermostability (up to 60°C) and specificity than M-MuLV Reverse Transcriptase

Packaging

1 kit containing 2 components

Quality

Absence of contaminants: Tested for the absence of detectable nonspecific RNases and nonspecific DNases in incubations with various nucleic acids (analyzed by gel electrophoresis) according to the current Quality Control procedures.
Function test: Reverse Transcriptase AMV is function tested in the cDNA Synthesis Kit and RT-PCR.

Unit Definition

1 unit is the enzyme activity that incorporates 1 nmol of [³H]-dTMP into acid-precipitable products in 10 minutes at +37 °C with poly(A)⁺ · d(pT)₁₅ as template primer.

Volume Activity: 20-25 U/μl. Please refer to the Certificate of Analysis for more information.

Preparation Note

Storage buffer: 200 mM potassium phosphate, 2 mM dithiothreitol, 0.2% Triton X-100 (v/v), 50% glycerol (v/v), pH 7.2.

Storage and Stability

Store at -15–-25 °C. (Storage at -70 °C is more likely to deteriorate the enzyme via repeated freezing and thawing.)

Other Notes

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

Kit Components Only

• Reverse Transcriptase AMV 20-25 U/μl
• First-strand cDNA Synthesis Buffer 5x concentrated

Safety Information

• Storage Class Code: 12 - Non Combustible Liquids
• WGK: WGK 1
• Flash Point(F): does not flash
• Flash Point(C): does not flash