Description

• Title:
  Electric chips for rapid detection and quantification of nucleic acids
• Creator:
  Gabig-Cimińska, Magdalena;
  Holmgren, Anders;
  Andresen, Heiko;
  Barken, Kim Bundvig;
  Wumpelmann, M;
  Albers, Joerg;
  Hintsche, Rainer;
  Breitenstein, Antje;
  Neubauer, Peter;
  Łoś, Marcin;
  Czyż, Agata;
  Węgrzyn, Grzegorz;
  Silfvesparre, Gustav;
  Jurgen, Britta;
  Schweder, Thomas;
  Enfors, Sven-Olof
• Description:
  A silicon chip-based electric detector coupled to bead-based sandwich hybridization (BBSH) is presented as an approach to perform rapid analysis of specific nucleic acids. A microfluidic platform incorporating paramagnetic beads with immobilized capture probes is used for the biorecognition steps. The protocol involves simultaneous sandwich hybridization of a single-stranded nucleic acid target with the capture probe on the beads and with a detection probe in the reaction solution, followed by enzyme labeling of the detection probe, enzymatic reaction, and finally, potentiometric measurement of the enzyme product at the chip surface. Anti-DIG–alkaline phosphatase conjugate was used for the enzyme labeling of the DIG-labeled detection probe. p-Aminophenol phosphate (pAPP) was used as a substrate. The enzyme reaction product, p-aminophenol (pAP), is oxidized at the anode of the chip to quinoneimine that is reduced back to pAP at the cathode. The cycling oxidation and reduction of these compounds result in a current producing a characteristic signal that can be related to the concentration of the analyte. The performance of the different steps in the assay was characterized using in vitro synthesized RNA oligonucleotides and then the instrument was used for analysis of 16S rRNA in Escherichia coli extract. The assay time depends on the sensitivity required. Artificial RNA target and 16S rRNA, in amounts ranging from 1011 to 1010 molecules, were assayed within 25 min and 4 h, respectively.
• Object availability:
  free access
• Date:
  2004
• Type:
  Article;
  PeerReviewed
• Language:
  English
• Format:
  application/pdf
• Publisher:
  Elsevier
• Relation: http://eprints.ibb.waw.pl/1144 ;http://www.sciencedirect.com/science/article/pii/S0956566303002732
• Subject:
  Q Science (General);
  QH Natural history;
  QH301 Biology;
  QH426 Genetics;
  T Technology (General)
• Identifier: http://eprints.ibb.waw.pl/1144/1/Preprint.pdf ;
  Gabig-Cimińska, Magdalena and Holmgren, Anders and Andresen, Heiko and Barken, Kim Bundvig and Wumpelmann, M and Albers, Joerg and Hintsche, Rainer and Breitenstein, Antje and Neubauer, Peter and Łoś, Marcin and Czyż, Agata and Węgrzyn, Grzegorz and Silfvesparre, Gustav and Jurgen, Britta and Schweder, Thomas and Enfors, Sven-Olof (2004) Electric chips for rapid detection and quantification of nucleic acids. Biosensors & bioelectronics, 19 (6). pp. 537-546. ISSN 1873-4235
• Data provider:
  Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk
• Can I use it?:
  seek permission
• Type:
  other