FISH & Fluorescent Reporters
Identifying the presence of targets in a sample can be challenging when working with out-dated, generic catalog probes that are not optimized to your specific project.
Let Daicel Arbor Biosciences help with sets of custom designed oligo probes to address specific research questions. With the wealth of genomic data available today, we can design unique pools of oligos specific to the molecular targets of interest. Improve binding efficiency with short oligos that easily penetrate cell barriers and maintain consistent hybridization profiles for simplified assays. We offer both immortal libraries that can be labeled in-house or labeled libraries that are ready-to-use upon arrival. Proprietary design software uses tried-and-true bioinformatic methods and the latest genomic assemblies, yielding in reduced background signal from repeat-rich regions and eliminating non-specific hybridization. Our team of experts provide complimentary assistance in design and selection of probes in order to deliver consistent, reliable results for any FISH project.
Fluorescent in situ Hybridization – FISH
Fluorescent in situ hybridization (FISH) allows visualization of any region of the genome using fluorescently labeled oligonucleotides. Traditionally these labeled oligonucleotide probes are generated from bacterial artificial chromosomes (BAC) which have been amplified with PCR and labeled using nick translation to incorporate fluorophores or haptens along the entire length of kilobase-sized probes. The size of these probes provides difficulties for efficient penetration of cell barriers, and they are additionally limited in selection from pre-designed catalogs of cloned genes, making them ill-suited to address advanced investigations into genomic structure.
myTags® are FISH probe libraries which are custom-designed using assembled sequences to address many of the drawbacks associated with BAC-derived probes. The FISH probes are designed with shorter hybridization sequences to allow for more efficient crossing of cell barriers. The proprietary bioinformatics design process at Daicel Arbor Biosciences uses custom genomic coordinates from assembled sequences for identification of hundreds to thousands of highly-specific probe sequences for hybridization to only intended targets. These probes collectively provide bright and reliable signal in FISH assays while minimizing or eliminating background signal associated with repetitive sequences and non-specific binding. Probe sequences are produced using our proprietary oligo synthesis technology and converted into FISH probes using the highly efficient myTags labeling method.
Detect the presence or absence of oligo sequence with custom designed myTags probes. myTags are short 5’ end labeled oligos that are designed to be highly-specific to intended oligo targets. Probes are synthesized in pools containing up to thousands of unique sequences, and labeled with fluorophores or haptens. These pools of probes hybridization to their intended targets and allow for the detection of the presence of targets through collective signal. Fluorescent probes can be used to enhance signals in microarray gene expression analyses, or in flow cytometry applications.
B. J. Beliveau, et al. (2012). Versatile design and synthesis platform for visualizing genomes with Oligopaint FISH probes. PNAS
B. J. Beliveau, et al. (2014). Visualizing Genomes with Oligopaint FISH Probes. Current Protocols in Molecular Biology
E. Joyce, et al. (2013). Germline Progenitors Escape the Widespread Phenomenon of Homolog Pairing during Drosophila Development. PLOS Genetics
Murgha et al. (2015) Combined in vitro transcription and reverse transcription method to amplify and label complex synthetic oligonucleotide probe libraries. Biotechniques
Murgha et al. (2014) Methods for the Preparation of Large Quantities of Complex Single-Stranded Oligonucleotide Libraries. PLoS ONE