Aptamer Taq HS (Glycerol-Free)
Aptamer Taq HS (glycerol-free) is a fast, high concentration, glycerol-free DNA polymerase containing a DNA aptamer that is designed to reversibly block the DNA polymerase and so work as a temperature-dependent hot start system. The glycerol-free formulation can be used to produce lyophilized or air-dried, ambient-temperature stable PCR or qPCR reagents and exhibit the same high performance before and after drying, offering greater flexibility for assay design and format.
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Compatible with highly sensitive reverse transcription
Four viral sequences, two DNA (Cytomegalovirus (green line) and Adenovirus (blue line)) and two RNA (Rotavirus (orange line) and Norovirus (red line)) were amplified using A/ Aptamer Taq HS (glycerol-free) (MDX015) or B/ Roche AptaTaq DNA Polymerase, with Lyo-Ready qPCR Buffer (MDX022) and RNase-Tolerant MMLV-RT (MDX043) in quadruplex qPCR probe assays. The data demonstrates both DNA and RNA templates are amplified with equal sensitivity using Aptamer Taq HS (glycerol-free), unlike the AptaTaq DNA Polymerase that gave very poor results with the RNA templates.
Traditional hot start PCR methods that are antibody or chemically mediated require a long activation time at very high temperatures. In contrast, DNA aptamers disassociate from the enzyme (e.g. activate) at relatively lower temperatures (45 °C), allowing for room temperature assay set-up, fast PCR reaction protocols and better assay specificity. Aptamer Taq HS (glycerol-free) is therefore suitable for difficult, multiplex PCR tests that require fast amplification whilst maintaining both high specificity and sensitivity.
|Description||Glycerol-free high concentration hot-start Taq DNA polymerase, suitable for difficult, multiplex molecular tests that require fast amplification, while maintaining both high specificity and sensitivity.|
|Concentration||Between 50 and 60 U/µL (see CoA)|
|Appearance||Clear, colorless solution|
|Hot Start||Reversible, temperature-dependent aptamer mediated|
|Application||PCR, real-time PCR, two-step RT-qPCR|
|Sample type||cDNA, DNA|
|Mix stability||See outer label|
|Assay stability||Up to 24 months at ambient temperature following air-drying or lyophilization|
|Purity||>50 % as measured as a percentage of total protein by quantitative gel
electrophoresis on Bioanalyzer (Agilent)
|DNA contamination||None detected in qPCR amplification with traces overlay with the negative control on E. coli and mouse genomic DNA specific targets.|
FAQs: Aptamer Taq HS (glycerol-free)
DNA aptamers are engineered oligonucleotides that bind to the active site of the DNA polymerase but are denatured and released as the temperature is increased during the PCR amplification cycle. This aptamer-mediated inhibition/activation process is however fully reversible, and so at the end of thermal cycling, when the temperature of the reaction is decreased, the DNA aptamer refolds and rebinds to DNA polymerase, inhibiting any further activity in the sample. This has proven to be important in workflows where undesired polymerase activity after reaction completion can disturb baseline readings, such as with high-throughput automated systems.
No, the DNA aptamer-based hot start PCR mechanism does not require an initial incubation. The aptamer is released from the DNA polymerase during normal cycling conditions, so Standard Taq protocols can be used.
No, the ends of the DNA aptamer are blocked to prevent them being used as primers during amplification.
Aptamer Taq HS (glycerol-free) is at 50 U/µL for PCR kit manufactures and DNA testing facilities, requiring large amounts of DNA polymerases. The highly concentrated Aptamer Taq HS (glycerol-free) allows for concentrated bulk production of PCR master mixes.
Concentrated bulks in glycerol-free formats are also attractive when using automated pipetting robots and other applications where accurate pipetting of small volumes is crucial, as glycerol makes the solution more viscous, the aptamer has the additional advantage of also allowing convenient room temperature reaction set-up.
Concentrated PCR master mixes are easier to lyophilize due to the reduced volume of the concentrated mix.
Glycerol is a cryoprotectant and is normally part of the storage buffer, where it serves to protect Taq DNA polymerase during freezing conditions. The high concentration of Aptamer Taq HS (glycerol-free) means that you get the same protection to freeze/thawing as a DNA polymerase that contains glycerol.
A lyophilized PCR molecular test will remain stable for up to 2 years at ambient temperature if correctly stored in sealed pouches.
Yes, Glycerol-Free Taq HS 50 U/µL is ideal for any multiplex PCR assays, particularly quantitative PCR molecular tests that require fast PCR amplification, whilst maintaining both high specificity and sensitivity.
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