Detail publikace

Pyrosequencing on a glass surface

ALMEIDA, A. MANZ, A. NEUŽIL, P.

Originální název

Pyrosequencing on a glass surface

Typ

článek v časopise ve Web of Science, Jimp

Jazyk

angličtina

Originální abstrakt

We demonstrate the use of open-surface microfluidics to sequence DNA by pyrosequencing at the plain hydrophobically coated surface of a microscope glass cover slip. This method offers significant advantages in terms of instrument size, simplicity, disposability, and functional integration, particularly when combined with the broad and flexible capabilities of open-surface microfluidics. The DNA was incubated on superparamagnetic particles and placed on a hydrophobically coated glass substrate. The particles with bound DNA were moved using magnetic force through microliter-sized droplets covered with mineral oil to prevent water evaporation from the droplets. These droplets served as reaction “stations” performing pyrosequencing as well as washing stations. The resequencing protocol with 34-mer single-stranded DNA (ssDNA) was used to determine the reaction performance. The de novo sequencing was performed with 51-mer and 81-mer ssDNA. The method can be integrated with previously shown sample preparation and PCR into a single sample-to-answer system on a plain glass surface.

Klíčová slova

DNA sequence determination; pyrosequencing; droplets; magnetic particles; open-surface microfluidics; virtual reaction chambers

Autoři

ALMEIDA, A.; MANZ, A.; NEUŽIL, P.

Vydáno

16. 2. 2016

Nakladatel

Royal Society of Chemistry

Místo

Londyn

ISSN

1473-0197

Periodikum

LAB ON A CHIP

Ročník

16

Číslo

6

Stát

Spojené království Velké Británie a Severního Irska

Strany od

1063

Strany do

1071

Strany počet

9

URL

https://pubs.rsc.org/en/content/articlelanding/2016/LC/C6LC00114A

Plný text v Digitální knihovně

http://hdl.handle.net/11012/203127

BibTex

@article{BUT122201,
  author="Ana V. {Almeida} and Andreas {Manz} and Pavel {Neužil}",
  title="Pyrosequencing on a glass surface",
  journal="LAB ON A CHIP",
  year="2016",
  volume="16",
  number="6",
  pages="1063--1071",
  doi="10.1039/c6lc00114a",
  issn="1473-0197",
  url="https://pubs.rsc.org/en/content/articlelanding/2016/LC/C6LC00114A"
}