DualHyb: Surface melting curve data from direct hybridization experiment of short
oligonucleotide probes with bimodal melting curve pattern.
Description
A melting curve experiment with four microbead populations and short
oligonucleotide probes (direct hybridization) and longer probes
(dual-hybridization probes) capture probe. Detection probes for human VIM
(vimentin), MLC-2v (myosin regulatory light chain 2, ventricular/cardiac
muscle isoform) and SERCA2 (Atp2a2 - ATPase, Calcium-transporting ATPase
sarcoplasmic reticulum type, slow twitch skeletal muscle isoform). One
sequence of VIM contained a mutation at position 41.
Arguments
Format
A data frame with the melting curves of three different capture and
detection probe pairs for HRPT1 and MLC-2v. First column contains the
temperature (in degree Celsius, 0.5 degree Celsius per step) followed by
melting curves of HRPT1 on 12 microbead populations and melting curves of
MLC-2v on 12 microbead populations.
T
a numeric
vector, Temperature in degree Celsius.
MLC2v
a numeric vector, MLC-2v with quencher-labeled
detection probe
SERCA2
a numeric vector, SERCA2 without quencher-labeled
detection probe
VIM.w.Mutation
a numeric vector, mutated VIM with
quencher-labeled detection probe
VIM.wo.Mutation
a numeric vector, native VIM with
quencher-labeled detection probe
Source
Data were measured with the VideoScan platform: A Highly Versatile Microscope Imaging Technology Platform for the Multiplex
Real-Time Detection of Biomolecules and Autoimmune Antibodies. S. Roediger,
P. Schierack, A. Boehm, J. Nitschke, I. Berger, U. Froemmel, C. Schmidt, M.
Ruhland, I. Schimke, D. Roggenbuck, W. Lehmann and C. Schroeder.
Advances in Biochemical Bioengineering/Biotechnology. 133:35--74,
2013. http://www.ncbi.nlm.nih.gov/pubmed/22437246 Surface Melting Curve Analysis with R. S. Roediger, A. Boehm and I.
Schimke. The R Journal. 5(2):37--52, 2013.
http://journal.r-project.org/ Nucleic acid detection based on the use of microbeads: a review. S.
Roediger, C. Liebsch, C. Schmidt, W. Lehmann, U. Resch-Genger, U. Schedler,
P. Schierack. Microchim Acta 2014:1--18. DOI:
10.1007/s00604-014-1243-4
Details
The melting curve was conducted with short oligonucleotide probes (direct
hybridization) and longer probes (dual-hybridization probes) on the surface
of microbeads (sequences and materials according to Roediger et al. (2012))
using the VideoScan platform by Roediger et al. (2012). The dyes and
quencher used were Atto 647N and BHQ2.