Tth111 Reverse Transcriptase

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Certificate of Analysis PDF -Current Lot
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Source

Thermus thermophilus

Reagents Supplied

10X Tth111 Reverse Transcriptase Reaction Buffer
MnCl₂ Solution 

Applications

• Suitable for high temperature synthesis of DNA (1)
• 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)

Unit Definition

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

Assay Conditions

40 mM Tris-HCl (pH 8.5)
1 mM MnCl₂
10 mM dithiothreitol
1 mg/ml bovine serum albumin
0.4mM 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

References

(1) Wang, A.M., Doyle, M.V. and Mark, D.F. (1989) Proc. Natl. Acad. Sci. USA 86, 9717-9721
(2) Myers, T.W., Gelfanld, D.H. (1991) Biochem. 30, 7661-7666
(3) Kather, H.L. and Schwartz, I. (1994) Biotechniques 16, 84-92
(4) Poddar, S.K., Sawyer, M.H. and Connor, J.D. (1998) J. Med. Microbiol. 47, 1131-1135