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@PHDTHESIS{Schilcher:822040,
      author       = {Schilcher, Felix Maximilian},
      othercontributors = {Weinhold, Elmar and Wöll, Dominik},
      title        = {{B}arcoding von genomischer {DNA} und {A}nwendung im
                      {B}ereich des {DNA}-{M}appings},
      school       = {RWTH Aachen University},
      type         = {Dissertation},
      address      = {Aachen},
      publisher    = {RWTH Aachen University},
      reportid     = {RWTH-2021-06512},
      pages        = {1 Online-Ressource : Illustrationen},
      year         = {2021},
      note         = {Veröffentlicht auf dem Publikationsserver der RWTH Aachen
                      University; Dissertation, RWTH Aachen University, 2021},
      abstract     = {In the course of this work, the sequence specific
                      modification of DNA using methyltransferase-directed
                      Transfer of Activated Groups (mTAG) was further developed.
                      One focus of this work was the synthesis of analogs of the
                      natural cofactor of DNA methyltransferases (DNA-MTases)
                      AdoMet. This includes AdoMet analogs for the transfer of
                      reactive as well as fluorescent and sterically demanding
                      groups. The latter includes analogs with
                      oligodeoxynucleotide modifications. In addition, the
                      enzymatic activity of the new AdoMet analogs was studied
                      using the DNA-MTases M.TaqI and M.BseCI. Compared to that of
                      reactive and fluorescent AdoMet analogs, the activity of
                      ODN-modified AdoMet analogs was generally lower and differed
                      greatly from one another. EMSA experiments were used to show
                      the influence of the sterically demanding modification on
                      electrophoretic mobility of modified DNA-fragments. On this
                      basis, one-step modification reactions were performed using
                      the newly developed AdoMet analogs. The modified DNA was
                      then analyzed using different methods, either optically or
                      electronically. This includes analysis using the so called
                      C-Trap as well as nanopores, which were carried out
                      externally by cooperation partners, as well as the analysis
                      of stretched and immobilized DNA using fluorescence
                      microscopy. Another focus of this work was the experimental
                      development of DNA modification using the mTAG method. On
                      one hand, new specificities were introduced implementing a
                      newly developed labelling strategy called DNA-blocking. On
                      the other hand, the mTAG method was extended to genomic DNA
                      embedded in agarose plugs. Finally, standardized
                      computational analysis routines were developed, which were
                      then used to automate the analysis of the fluorescence
                      microscopy of stretched and immobilized DNA to determine and
                      compare the quality of different modification experiments on
                      the basis of key figures.},
      cin          = {152620 / 150000},
      ddc          = {540},
      cid          = {$I:(DE-82)152620_20140620$ / $I:(DE-82)150000_20140620$},
      typ          = {PUB:(DE-HGF)11},
      doi          = {10.18154/RWTH-2021-06512},
      url          = {https://publications.rwth-aachen.de/record/822040},
}