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CAL Fluor® Gold 540 dye fluoresces in the yellow green region of the visible spectrum. CAL Fluor Gold 540 amidite is used for the 5' labeling of fluorogenic probes used in 5' nuclease assays, Molecular Beacons, Scorpions® primers and other detection assays. This amidite does not contain a DMT protecting group and can only be added to the 5' terminus of the oligo. CAL Fluor Gold 540 fluorophore is an alternative for TET and is quenched by BHQ®-1.
Properties:
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Chemical Name:
Diisopropyl-phosphoramidous acid 2-cyano-ethyl ester 1-[2-(6-ethylamino-2, 7-dimethyl-3-oxo-3H-xanthen-9-yl)-benzoyl]-piperidin-4-yl ester
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Formula:
C38H47N4O5P
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Molecular Weight:
670.33
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Appearance:
orange-red solid
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Absorption Maximum (Lambda Max):
522
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Extinction Coefficient at Lambda max:
81100
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Extinction Coefficient at 260 nm:
15100
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Fluorescence Maximum:
543
Product Usage:
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Synthesis Conditions:
To prevent decomposition of CAL Fluor Gold, we recommend using CAL Fluor Gold with fast deprotecting amidites and mild deprotection conditions. Dilute to the recommended concentration, mix thoroughly in sealed vial and allow to sit for 3 minutes to ensure that all contents are dissolved in DCM. We recommend using Ethyl thiol tetrazole for the activator reagent with this product.
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Dilution:
100 µmol/mL
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Coupling:
15-minute coupling
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Deprotection Conditions:
Cleave and deprotect the CAL dye labeled oligo, from the CPG support, using either 2-methoxyethylamine (Ald# 241067-50) and MeOH (1:3) for 3 hours at 60 °C or t-butylamine/water (1:3) for 6 hours at 60 °C. Filter sample and evaporate supernatant. Re-dissolve the CAL dye labeled oligo in water. The use of strong bases, such as concentrated ammonia may cause degradation of the dye and should be avoided.
Cleave and deprotect the CAL dye labeled oligo, from the CPG support, using either 2-methoxyethylamine (Ald# 241067-50) and MeOH (1:3) for 3 hours at 60 °C or t-butylamine/water (1:3) for 6 hours at 60 °C. Filter sample and evaporate supernatant. Re-dissolve the CAL dye labeled oligo in water.
The use of strong bases, such as concentrated ammonia may cause degradation of the dye and should be avoided.
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The mass this product adds after conjugation and work-up (the additional mass seen by mass spectrometry) is:
532.52
Image of cleaved and deprotected structure:
Storage and Handling:
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Shipping Conditions:
Cold
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Storage Conditions:
-15 to -30 °C
