Experimental Protocol
TthPlus-Taq DNA Taq DNA Polymerase
TthPlus-Taq DNA Polymerase
Synonyms
TthPlus-Taq DNA; TthPlus-Taq DNA Polymerase; TthPlus DNA Polymerase; TthPlus-Taq DNA taq dna polymerase
Form/Format
10 mM K-phosphate buffer pH 7.0 (25°C), 100 mM NaCl, 0.5 mM EDTA, 1 mM DTT, 50% glycerol (v/v), 0,1 mg/ml BSA.
Concentration
5 units/ul (varies by lot)
Application Notes
- Taq enzyme having both reverse transcription and DNA amplification activity, excellent for one step RT-PCR and Real-Time Quantitative PCR;
- For high temperature reverse transcription that allows highly specific priming of cDNA synthesis and overcomes problems with secondary structure.
- For high temperature reverse transcription that allows highly specific priming of cDNA synthesis and overcomes problems with secondary structure.
Features
• thermostable
• DNA polymerase activity in the presence of MgCl2
• reverse transcriptase activity in the presence of MnCl2
• reverse transcription at elevated temperature minimizing secondary structure problems
• DNA polymerase activity in the presence of MgCl2
• reverse transcriptase activity in the presence of MnCl2
• reverse transcription at elevated temperature minimizing secondary structure problems
Unit Definition
One unit is defined as the amount of enzyme that incorporates 10 nmoles of dNTPs into acid-insoluble form in 30 minutes at 72°C under the following reaction conditions: 25 mM TAPS buffer (Tris-(hydroxymethyl)-methyl- amino-propanesulfonic acid, sodium salt) pH 9.3 (25°C), 50 mM KCl, 2 mM MgCl2, 1 mM β-mercaptoethanol, 200 uM dNTPs and 10 ug of calf thymus DNA in a final reaction volume of 50 ul.
Component
- TthPlus DNA polymerase, 500 units
- 10 x OneTube buffer, 1 ml
- 50 mM Mn(OAc)2, 0.75ml
Note: The optimal experimental conditions depend on the system used and they should be individually determined. The Mn2+ concentrations and the enzyme amount are the limiting factors for an accurate result. Traditionally 5 units of enzyme and a Mn-acetate concentration of 3.5 mM are used for the reverse transcription in a final 50 ul reaction volume.
- 10 x OneTube buffer, 1 ml
- 50 mM Mn(OAc)2, 0.75ml
Note: The optimal experimental conditions depend on the system used and they should be individually determined. The Mn2+ concentrations and the enzyme amount are the limiting factors for an accurate result. Traditionally 5 units of enzyme and a Mn-acetate concentration of 3.5 mM are used for the reverse transcription in a final 50 ul reaction volume.
Preparation and Storage
Store TthPlus DNA polymerase preferably at -20°C in a constant temperature freezer.
Experimental Protocol
Experimental Protocol
Related Product Information for TthPlus-Taq DNA taq dna polymerase
TthPlus DNA polymerase is isolated from the Thermus thermophilus strain. TthPlus DNA polymerase is a single 92 kDa polypeptide showing a 5'-3' exonuclease activity but lacking 3'-5' exonuclease activity. It catalyzes the polymerization of nucleotides into double-stranded DNA in the presence of MgCl2. Its efficiency has been shown more particularly on large DNA fragments up to 12 kb (using lambda phage DNA as a template). TthPlus DNA polymerase is also capable of catalyzing the polymerization of DNA using a RNA template in the presence of Mn2+. The ability of TthPlus DNA polymerase to reverse transcribe at elevated temperatures (70°C) minimizes the problems encountered with strong secondary structures in RNA since they are unstable at higher reaction temperatures. Higher temperatures also result in increased specificity of primer hybridization and extension. In coupled RT/PCR assays, TthPlus is about 50-100 times more efficient than regular Taq DNA polymerase.
TthPlus DNA polymerase is delivered with a one-tube 10x buffer. Both reactions (RT and PCR) are carried out in the same buffer and same tube with the same enzyme (TthPlus). The 10 x one tube buffer does not contain Mn(OAc). Mn(OAc) is provided extra and have to be added at a final concentration of 3.5 mM.
TthPlus DNA polymerase is delivered with a one-tube 10x buffer. Both reactions (RT and PCR) are carried out in the same buffer and same tube with the same enzyme (TthPlus). The 10 x one tube buffer does not contain Mn(OAc). Mn(OAc) is provided extra and have to be added at a final concentration of 3.5 mM.
Product Categories/Family for TthPlus-Taq DNA taq dna polymerase
Similar Products
Product Notes
The TthPlus-Taq DNA (Catalog #AAA433681) is a Taq DNA Polymerase and is intended for research purposes only. The product is available for immediate purchase. - Taq enzyme having both reverse transcription and DNA amplification activity, excellent for one step RT-PCR and Real-Time Quantitative PCR; - For high temperature reverse transcription that allows highly specific priming of cDNA synthesis and overcomes problems with secondary structure. Researchers should empirically determine the suitability of the TthPlus-Taq DNA for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "TthPlus-Taq DNA, Taq DNA Polymerase" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
If you are ready to order, navigate to Shopping Cart and get ready to checkout.
