Merck
  • Home
  • Search Results
  • Effect of non-specific species competition from total RNA on the static mode hybridization response of nanomechanical assays of oligonucleotides.

Effect of non-specific species competition from total RNA on the static mode hybridization response of nanomechanical assays of oligonucleotides.

Nanotechnology (2014-05-09)
Rohit Mishra, Martin Hegner
ABSTRACT

We investigate here the nanomechanical response of microcantilever sensors in real-time for detecting a range of ultra-low concentrations of oligonucleotides in a complex background of total cellular RNA extracts from cell lines without labeling or amplification. Cantilever sensor arrays were functionalized with probe single stranded DNA (ssDNA) and reference ssDNA to obtain a differential signal. They were then exposed to complementary target ssDNA strands that were spiked in a fragmented total cellular RNA background in biologically relevant concentrations so as to provide clinically significant analysis. We present a model for prediction of the sensor behavior in competitive backgrounds with parameters that are indicators of the change in nanomechanical response with variation in the target and background concentration. For nanomechanical assays to compete with current technologies it is essential to comprehend such responses with eventual impact on areas like understanding non-coding RNA pharmacokinetics, nucleic acid biomarker assays and miRNA quantification for disease monitoring and diagnosis to mention a few. Additionally, we also achieved a femtomolar sensitivity limit for online oligonucleotide detection in a non-competitive environment with these sensors.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
丙酮, ACS reagent, ≥99.5%
Sigma-Aldrich
丙酮, suitable for HPLC, ≥99.9%
Sigma-Aldrich
二乙醚, contains 1 ppm BHT as inhibitor, anhydrous, ≥99.7%
Sigma-Aldrich
丙酮, HPLC Plus, for HPLC, GC, and residue analysis, ≥99.9%
Sigma-Aldrich
尿素, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
二乙醚, anhydrous, ACS reagent, ≥99.0%, contains BHT as inhibitor
Sigma-Aldrich
尿素, ACS reagent, 99.0-100.5%
Sigma-Aldrich
二乙醚, suitable for HPLC, ≥99.9%, inhibitor-free
Sigma-Aldrich
二乙醚, ACS reagent, anhydrous, ≥99.0%, contains BHT as inhibitor
Sigma-Aldrich
丙酮, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.5% (GC)
Sigma-Aldrich
二乙醚, contains BHT as inhibitor, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., ≥99.8%
Sigma-Aldrich
尿素, ReagentPlus®, ≥99.5%, pellets
Sigma-Aldrich
丙酮, suitable for HPLC, ≥99.8%
Sigma-Aldrich
丙酮, Laboratory Reagent, ≥99.5%
Sigma-Aldrich
尿素 溶液, BioUltra, ~8 M in H2O
Supelco
丙酮, analytical standard
Supelco
尿素, 8 M (after reconstitution with 16 mL high purity water)
Sigma-Aldrich
尿素, BioXtra, pH 7.5-9.5 (20 °C, 5 M in H2O)
Sigma-Aldrich
尿素, suitable for electrophoresis
Sigma-Aldrich
尿素, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.0-100.5%, 99.0-101.0% (calc. on dry substance)
Supelco
丙酮, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
尿素, BioUltra, for molecular biology, ≥99% (T)
Sigma-Aldrich
丙酮, ACS reagent, ≥99.5%
Sigma-Aldrich
尿素, puriss. p.a., ACS reagent, reag. Ph. Eur., ≥99%
Sigma-Aldrich
丙酮, suitable for HPLC, ≥99.9%
USP
丙酮, United States Pharmacopeia (USP) Reference Standard
Sigma-Aldrich
二乙醚, ACS reagent, ≥98.0%, contains ≤2% ethanol and ≤10ppm BHT as inhibitor
Supelco
二乙醚, analytical standard
Sigma-Aldrich
丙酮, histological grade, ≥99.5%
Sigma-Aldrich
丙酮, natural, ≥97%