Lambda exonuclease pre-treatment for improved DNA-chip performance depends on the relative probe-target position.
The hybridisation characteristics of DNA targets to solid phase bound probes, e.g. in DNA microarrays, depend on the probe-target position and on target renaturation if a dsDNA target is used. We investigated a lambda exonuclease treatment of a PCR amplified dsDNA target to produce ssDNA with regard to probe-target position, treatment duration and inactivation time towards its impact on fluorescence or electrical signals on two DNA-chip formats. Surprisingly, the achieved amplification factors varied by three orders of magnitude, i.e. 2-1074 fold signal enhancement, depending on the relative probe-target position and readout scheme. The presented results can be used to design future studies involving lambda exonuclease preanalytic treatments.
Brinker
Anja
A
Jenaer Biochip Initiative, Friedrich Schiller University, Institute of Physical Chemistry, Helmholtzweg 4, Jena, Germany.
Schulze
Holger
H
Bachmann
Till
T
Möller
Robert
R
D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005090:Exodeoxyribonucleases / Q000737:chemistry* / Q000235:genetics*; D018390:Gene Targeting; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation*
D015342:DNA Probes; D005090:Exodeoxyribonucleases