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978-3-8439-2041-4, Reihe Organische Chemie

Sebastian Spies
Mehrfache Chemische Primerverlängerung am 5'-Terminus eines Aminoterminalen Primers

145 Seiten, Dissertation Universität Stuttgart (2014), Hardcover, A5

Zusammenfassung / Abstract

In the cell, both replication and transcription of genetic information occurs via primer extension in 5ʹ- to 3ʹ-direction. The goal of this work was to demonstrate, that the non-enzymatic copying of a DNA sequence is feasible via template-direct primer extension in the unnatural 3ʹ → 5ʹ-direction. To this end, a 5ʹ-amino-terminal DNA-primer was elongated in multiple primer extension reactions via reaction with activated nucleotide building blocks. First, a method was established for multiple primer extension on templates immobilized via hybridization to a capture oligonucleotide that had been reacted with epoxy modified magnetic beads. Products were read-out after every elongation step via MALDI TOF mass spectrometry. Secondly, active esters of nucleotides with a protected 5ʹ-amino group were synthesized.