title: On-chip laser-induced DNA dehybridization.
abstract: Detection of pathogens and relevant genetic markers using their nucleic acid signatures is extremely common due to the inherent specificity genomic sequences provide. One approach for assaying a sample simultaneously for many different targets is the DNA microarray, which consists of several million short nucleic acid sequences (probes) bound to an inexpensive transparent substrate. Typically, complex samples hybridize to the microarray and the pattern of fluorescing probes on the microarray's surface identifies the detected targets. In the case of evolving or newly emergent organisms, a hybridization pattern can occur that differs from any previously known sources. When this happens it can be useful to recover the hybridized DNA from the binding locations of interest for sequencing. Here we present the novel utilization of a focused Infrared (IR) laser to heat user-selected spots on the DNA microarray surface, causing only localized dehybridization and recovery of the desired DNA into an elution buffer where it is available for subsequent amplification or sequencing. The introduction of a focused dehybridization method for spots of interest suppresses the amount of background DNA to be analyzed from downstream processes, and should reduce subsequent sequence assembly errors. This technique could also be applied to high-density protein microarrays where the desire to locally heat spots for release of bound molecules is desired.
authors.affiliation: Center for Micro and Nano Technology, LLNL, Livermore, California, USA.
authors.forename: E K
authors.initials: EK
authors.lastname: Wheeler
authors.forename: B R
authors.initials: BR
authors.lastname: Baker
authors.forename: W T
authors.initials: WT
authors.lastname: Piggott
authors.forename: S L
authors.initials: SL
authors.lastname: Mabery
authors.forename: C A
authors.initials: CA
authors.lastname: Hara
authors.forename: J
authors.initials: J
authors.lastname: DeOtte
authors.forename: W
authors.initials: W
authors.lastname: Benett
authors.forename: E V
authors.initials: EV
authors.lastname: Mukerjee
authors.forename: J
authors.initials: J
authors.lastname: Dzenitis
authors.forename: N R
authors.initials: NR
authors.lastname: Beer
mesh_terms: D002138:Calibration; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D007834:Lasers; D046210:Microfluidic Analytical Techniques; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics
chemical_list: D015345:Oligonucleotide Probes; D012367:RNA, Viral; D004247:DNA