Single-molecule denaturation mapping of DNA in nanofluidic channels

Here we explore the potential power of denaturation mapping as a single-molecule technique. By partially denaturing YOYO (R)-1-labeled DNA in nanofluidic channels with a combination of formamide and local heating, we obtain a sequence-dependent "barcode" corresponding to a series of local dips and peaks in the intensity trace along the extended molecule. We demonstrate that this structure arises from the physics of local denaturation: statistical mechanical calculations of sequence-dependent melting probability can predict the barcode to be observed experimentally for a given sequence. Consequently, the technique is sensitive to sequence variation without requiring enzymatic labeling or a restriction step. This technique may serve as the basis for a new mapping technology ideally suited for investigating the long-range structure of entire genomes extracted from single cells.

Larsen, Niels B. (författare)
Silahtaroglu, Asli (författare)
Kristensen, Anders (författare)
Tommerup, Niels (författare)
Tegenfeldt, JonasLund University,Lunds universitet,Fasta tillståndets fysik,Fysiska institutionen,Institutioner vid LTH,Lunds Tekniska Högskola,Solid State Physics,Department of Physics,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)ftf-jte (författare)
Flyvbjerg, Henrik (författare)
Fasta tillståndets fysikFysiska institutionen (creator_code:org_t)

Ingår i:Proceedings of the National Academy of Sciences: Proceedings of the National Academy of Sciences107:30, s. 13294-132991091-64900027-8424

Av författaren/redakt...: Reisner, Walter; Larsen, Niels B.; Silahtaroglu, As ...; Kristensen, Ande ...; Tommerup, Niels; Tegenfeldt, Jona ...; visa fler...; Flyvbjerg, Henri ...; visa färre...

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