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Updated: Jul 5, 2026

Analysis of Termination of Transcription Using BrUTP-strand-specific Transcription Run-on (TRO) Approach
Published on: March 12, 2017
Protein truncation test
Rolf Vossen1, Johan T den Dunnen
1Human and Clinical Genetics, Leiden University Medical Center, Leiden, The Netherlands.
Abstract:
The protein truncation test detects mutations at the protein level that lead to premature translation termination. The method has evolved considerably since is original publication in this manual. This thoroughly revised unit describes what is now the preferred method for performing the protein truncation test. Transcription and translation are performed in separate reactions; during translation, biotin-labeled or N-terminally tagged proteins are synthesized. The translation products are detected on immunoblots via chemiluminescence. An Alternate Protocol using coupled in vitro transcription/translation and radiolabeled proteins is also presented.
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