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Single molecule transcription profiling with AFM*

Jason Reed1,6, Bud Mishra2, Bede Pittenger3, Sergei Magonov3, Joshua Troke4, Michael A Teitell4,5 and James K Gimzewski1,5,6

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Established techniques for global gene expression profiling, such as microarrays, face fundamental sensitivity constraints. Due to greatly increasing interest in examining minute samples from micro-dissected tissues, including single cells, unorthodox approaches, including molecular nanotechnologies, are being explored in this application. Here, we examine the use of single molecule, ordered restriction mapping, combined with AFM, to measure gene transcription levels from very low abundance samples. We frame the problem mathematically, using coding theory, and present an analysis of the critical error sources that may serve as a guide to designing future studies. We follow with experiments detailing the construction of high density, single molecule, ordered restriction maps from plasmids and from cDNA molecules, using two different enzymes, a result not previously reported. We discuss these results in the context of our calculations.


Footnote
*  Based on invited talk at the International Conference on Nanoscience and Technology 2006.
PACS

87.15.N- Properties of solutions of macromolecules

87.14.G- Nucleic acids

82.37.Rs Single molecule manipulation of proteins and other biological molecules

87.15.A- Theory, modeling, and computer simulation

Subjects

Biological physics

Nanoscale science and low-D systems

Chemical physics and physical chemistry

Dates

Issue 4 (31 January 2007)

Received 1 November 2006, in final form 5 December 2006

Published 21 December 2006



  1. Single molecule transcription profiling with AFM

    Jason Reed et al 2007 Nanotechnology 18 044032

  2. Dielectrophoretic reconfiguration of nanowire interconnects

    Alexander D Wissner-Gross 2006 Nanotechnology 17 4986

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