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Abstract for TNT® T7 Quick for PCR DNA
The TNT® T7 Quick for PCR DNA is a rapid,
convenient, coupled transcription/translation system designed for optimum
expression of PCR templates. For most PCR templates, the TNT®
T7 Quick for PCR DNA reactions produce up to 5 times more protein than other
commercially available kits. The PCR-generated DNA can be used directly from the
amplification reaction. PCR products from amplification protocols and commercially
available systems (e.g., Promega Access RT-PCR System [Cat.#
A1250], Roche Diagnostics High Fidelity and Expand™ Long
Template PCR Systems, and Invitrogen Platinum Taq) have been successfully tested. The system
will work with any other comparable PCR amplification scheme. The PCR products
may be added directly to the TNT® T7 Quick for PCR
DNA reaction without purification.
To use TNT® T7 Quick for PCR DNA, a PCR
fragment containing a T7 promoter is added to the TNT®
T7 PCR Quick Master Mix and incubated for 60–90 minutes at 30°C. The
synthesized proteins then can be analyzed by SDS-polyacrylamide gel
electrophoresis (SDS-PAGE) followed by autoradiography or phosphorimaging.
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