Biosensors & Bioelectronics

William Richard McCombie

Publications

• Research Article
  Fragmentation of Surface Adsorbed and Aligned DNA Molecules using Soft Lithography for Next-Generation Sequencing
  Author(s): NaHyun Cho, Sara Goodwin, Julia Budassi, Ke Zhu, William Richard McCombie and Jonathan SokolovNaHyun Cho, Sara Goodwin, Julia Budassi, Ke Zhu, William Richard McCombie and Jonathan Sokolov
  
  In this study, the enzymatic in situ cutting of linearized DNA molecules at approximately 11 kbp intervals is demonstrated using a soft lithography technique. The ultimate goal is to provide a general ordered cutting method to greatly simplify the assembly process. DNA was stretched onto PMMA (Poly methyl methacrylate) coated silicon by withdrawing the substrate from a DNA solution (a process termed “combing”). The stretched lambda DNA could be linearly cut with a soft lithography stamp used to selectively apply DNase I. After cutting the DNA on the substrate, the DNA fragments are removed from the surface by incubating PMMA in the commercial NEBuffer 3.1 at 75°C. The recovered fragments desorbed into the buffer and were sequenced using the PacBio RS II sequencer without an amplification step. The mean coverage was 2870X for the approximately 11 kbp fragmented sample a.. Read More»
  DOI: 10.4172/2155-6210.1000247

  Abstract PDF