Tth111 Reverse Transcriptase

Source

Thermus thermophilus 111

Applications

• Suitable for high temperature synthesis of DNA (1)
• Tth III Reverse Transcriptase synthesizes cDNA from RNA template (2)
• Results in greater specificity of primer hybridization and extension of RNA
• Can reverse transcribe at elevated temperatures (2)
• Minimizes problems with strong secondary structure of RNA
• Used for efficient PCR of DNA, containing problematic secondary structures
• Applicable to RT-PCR; the same enzyme is used for both reverse transcription and following amplification of obtained cDNA template (2)
• Resistant to amplification inhibitors present in template DNA isolated from problematic samples (3,4)

Reagents Supplied

10X Tth 111 Reverse Transcriptase Reaction Buffer
MnCl₂ Solution 

Unit Definition

One unit is the amount of enzyme required to incorporate 1.0 nmole TTP into acid insoluble material in 10 minutes at 50°C

Assay Conditions

40 mM Tris-HCl (pH 8.5)
1 mM MnCl₂
10 mM dithiothreitol
1 mg/ml bovine serum albumin
0.4 mM polyA(dT)
0.5 mM (α-³²P)TTP @ 10 μCi/ml 
Incubation is for 10 minutes at 50°C in a total reaction volume of 50 µl

Storage Buffer

50 mM Tris-HCl (pH 7.5)
0.1 mM EDTA
5.0 mM dithiothreitol
50% (v/v) glycerol
Stabilizers

Storage Conditions

Store at -20°C
Product shipped on Dry Ice