AMV Reverse Transcriptase

High quality enzyme for cDNA synthesis

Description

The AMV Reverse Transcriptase (AMV RT), is isolated from Avian Myeloblastosis Virus as the beta holoenzyme of molecular weight 157,000 daltons, using a modification of the method described by Houts et al. (1979). AMV RT is qualified for cDNA synthesis and also for dideoxy sequencing of DNA and RNA. AMV RT is compatible with 4 mM sodium pyrophosphate and placental RNase inhibitor in cDNA reactions. Using AMV Reverse Transcriptase, the reverse transcription step can be carried out at elevated temperatures up to 70 °C. High temperature increases the specificity of priming and helps to increase the efficiency by reducing RNA secondary structures. AMV RT is supplied with optimized 10x AMV Buffer.

Applications

• First-strand cDNA synthesis
• Dideoxy sequencing of DNA and RNA

Advantages

• Compatible with 4 mM sodium pyrophosphate and placental RNase inhibitor in cDNA reactions

Components

AMV RT is supplied in storage buffer: 0.2 M KPO₄ (pH 7.2), 2.0 mM DTT, 0.2 % Triton® X-100 and 50 % Glycerol (v/v), and provided with 10x AMV Buffer concentrate which is used as 10x or 5x stock depending on the application.

Reaction buffer concentrate
250 mM Tris-HCl (pH 8.3 at 25 °C), 50 mM MgCl₂, 500 mM KCl and 20 mM DTT. Thaw at 37 °C. Triturate with pipet tip to dissolve, if necessary.

Dilution buffer
10 mM KPO₄ (pH 7.2), 2.0 mM DTT, 0.2 % Triton X-100, 10 % Glycerol (v/v).

Storage and stability

Stable for 1 year from the assay date. Recommended storage temperature –20 °C (–70 °C for long term storage).
Note: Upon receipt, centrifuge the vial in a microfuge to ensure full recovery of the enzyme. In order to conserve optimal activity, it is strongly recommended that the enzyme be aliquoted and stored at -70 °C, although storage at -20 °C is adequate for short periods of time. Repeated freezing and thawing results in loss of enzyme activity. Due to the viscosity of the enzyme, maximum dispensing efficiency is achieved by the use of positive displacement pipets.

Ordering information

AMV Reverse Transcriptase	Size	Concentration
F-570S	500	20 U/µl
F-570L	2,500	20 U/µl