Description

• Title:
  Effect of unlabeled helper probes on detection of an RNA target by bead-based sandwich hybridization
• Creator:
  Barken, Kim Bundvig;
  Gabig-Cimińska, Magdalena;
  Holmgren, Anders;
  Molin, Søren
• Description:
  Unlabeled helper oligonucleotides assisting a bead-based sandwich hybridization assay were tested for the optimal placement of the capture and detection probes. The target used was a full-length in vitro synthesized mRNA molecule. Helper probes complementary to regions adjacent to the binding site of the 5' end attached capture probe were found much more effective than helper probes targeting positions adjacent to the detection probe binding site. The difference is believed to be caused by a disruption of the RNA secondary structure in the area where the capture probe binds, thereby reducing structural interference from the bead. The use of additional helpers showed an additive effect. Using helpers at both sides of the capture and detection probes showed a 15- to 40-fold increase in hybridization efficiency depending on the target, thereby increasing the sensitivity of the hybridization assays. Using an electrical chip linked to the detection probe for the detection of p-aminophenol, which is produced by alkaline phosphatase, a detection limit of 2 x 10(-13) M mRNA molecules was reached without the use of a nucleic acid amplification step.
• Object availability:
  free access
• Date:
  2004
• Type:
  Article;
  PeerReviewed
• Language:
  English;
  eng
• Format:
  application/pdf
• Publisher:
  Eaton Publishing
• Relation: http://eprints.ibb.waw.pl/1071 ;http://www.biotechniques.com/archive
• Subject:
  QH301 Biology;
  QH426 Genetics
• Identifier: http://eprints.ibb.waw.pl/1071/1/BTN_A_04361RR03_O_36123a.pdf ;
  Barken, Kim Bundvig and Gabig-Cimińska, Magdalena and Holmgren, Anders and Molin, Søren (2004) Effect of unlabeled helper probes on detection of an RNA target by bead-based sandwich hybridization. Biotechniques, 36 (1). pp. 124-132. ISSN 0736-6205
• Data provider:
  Repozytorium Instytutu Biochemii i Biofizyki Polskiej Akademii Nauk
• Can I use it?:
  seek permission
• Type:
  other