Enhanced Diagnostic Tools
Bio-Synthesis offers custom LI-COR IRDyes oligo labeling services. The IRDye family delivers near-infrared fluorescent imaging with enhanced sensitivity due to low background autoflourescence in the near-infrared region, and therefore, delivers high signal to noise ratios. IRDye fluorescent dyes have absorption and emission wavelengths in the NIR spectrum between 680 and 800nm.
The LI-COR IRDye family is optimized to be used with the LI-COR Infrared Imaging Systems, however the dyes can be used with a variety of instruments that offer near-infrared excitation and emission.
Advantages of IRDye near-infrared dyes:
IRDye labels can be incorporated at any position within an oligonucleotide at time of synthesis using phosphoramdite reagent for labeling. However certain modifications require the use of post-synthetic conjugation where a primary amine modified oligo is used to covalently attach with dye via NHS ester chemistry; other cross-linking chemistries are also available depending on particular project specifications. Single HPLC purification is recommended when use of phosphoramidite chemistry, dual HPLC is require for post-synthetic labeling.
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Our fluorescently labeled probes are manufactured using a rigorous process with purity of >95% pure for your high demand detection application.
Li-Cor dye can be incorporated into an oligonucleotide at the time of synthesis using phosphoramdite chemistry or conjugate post-synthetically using manual labeling of an activated dye with amino, thiol or other fucntional groups of the oligonucleotide.
Example of modifications using activated esters (NHS
ester) are coupled with amino groups whereas
thiol reactive dye reagents, like maleimides, are
coupled with thiol groups. In the first production
step, an amino- or thiol-modified oligonucleotide
is synthesised, desalted and purified by HPLC, the chemical
reaction between linking group and activated dye
is carried out, validated and purified by HPLC to remove unreact molecules.
Li-COR IRDye labels such as
IRDye 650 and quencher,
IRDye QC-1 Infrared Dye all require post-synthetic labeling. Order of such require dual HPLC purification.
All our oligonucleotide undergo vigorous process monitoring and strict quality control. Length and labeling are systematically controlled by PAGE and characterized by MALDI-TOP mass spectrometry analysis. Quantity is determined and validated by Fluoroscan (ABS/EM).
You will receive a Quality Assurance Certificate that includes quantitated yield, melting temperature, MW and µg/OD with every oligo. You will also receive an additional label for use in your records or on an additional vial.
Desalt or cartridge (RP1) purification is acceptable for most common phosphoramidite modifiers. However, double purification with HPLC is strongly recommended for modifiers that require NHS-ester chemistry to conjugate a dye through an amine linker such as Molecular Probes dyes. Failure sequences are removed during the first HPLC; the second HPLC is performed to remove unlabeled oligonucleotides and excessive dye.
Expected shipment time for common dye modifiers is 3 - 5 working days. More exotic modifications are usually completed within 5 - 7 working days.
680 and 800nm IRDye infrared Dyes are compatible with the Odyssey® CLx, Odyssey Fc, Odyssey Sa, Pearl® imager, Pearl Impulse imager, and 4300 DNA Analyzer platforms.
*IRDye 750 and IRDye 650 are not optimal for use on LI-COR imaging systems