%PDF-1.6 %���� 0000026076 00000 n Visit us on the Web at discover.bio-rad.com Bio-Rad Laboratories, Inc. Life Science Group 2000 Alfred Nobel Drive Hercules, CA 94547 USA For example, FAM is a good dye choice to detect low copy transcripts because it has high uorescent signal intensity. 0000051527 00000 n For example, FAM is a good dye choice to detect low-copy transcripts because it has high fluorescent signal intensity. Wavelength scan from 480 nm to 700 nm with excitation at 450 nm. wavelength of absorption and the quantitative issue of the inten-sity of absorption. As long as the two dyes are close to each other (within 15 nucleotides), the energy emitted by the donor dye excites the acceptor dye on the second HybProbe probe, which then emits fluorescent light at a different wavelength. 0000018031 00000 n FAM Emission. This structure features a linker between Coumarin 343 fluorophore, and the functional group. Consider total uorescence intensity as well. 0000001717 00000 n 0000017395 00000 n trailer TET. Spectral Properties . 0000049106 00000 n The photogenerated electrons of FAM were then transferred to Zr ions with a high-electron-accepting ability of Zr–O clusters in PCN-222, leading to fluorescence quenching toward FAM. 0000001164 00000 n 0000017792 00000 n MP = Mustang Purple™ dye. The principles of fluorescence and the synthesis of fluorescent oligonucleotides are described below. 0000026876 00000 n We have developed the new It provides better solubility, and spatial separation between the dye, and the functional group. 0000069565 00000 n 118 0 obj <>stream TAMRA Absorbance. Lifetime Data of Selected Fluorophores. Rhodamine Green-X. 76 0 obj <> endobj H���Y�%������n��Έ�A<3B���AƘq���Hh4����D՝i/�з"+32�'�����O߿~z�>����_�J%}��W/���K�^��m�"�cߔ�j���^�|[ҫ_.��������^����/גK����·ߓy�J3�}�. 0000042483 00000 n startxref 0000027051 00000 n 519. Wavelength (nm) FAM Absorption and Emission Spectra . 520. 0000002599 00000 n Below pH 7, 6-FAM™ becomes protonated, which results in decreased fluorescence. A single dye is excited at a particular wavelength and emits a photon at a higher wavelength. 0000010927 00000 n Choose dyes with appropriate excitation wavelengths and little to no overlap in their emission spectra. FAM Absorbance. <]/Prev 83581/XRefStm 1404>> H��V�v�0��+��v�=�Rާ���9�E�����@�����r�� You can then use 0000014285 00000 n 0000048744 00000 n 0000025302 00000 n Coumarin 343 is a blue emitting dye that can serve as a FRET donor for fluorescein (FAM). 0000030517 00000 n This can be accomplished using non-specific DNA binding dyes or sequence-specific probes. 0000069316 00000 n 0000045848 00000 n 0000001616 00000 n four reporter dyes, a new quencher, and a new master mix. endstream endobj 117 0 obj <>/Filter/FlateDecode/Index[8 68]/Length 21/Size 76/Type/XRef/W[1 1 1]>>stream lists many of the fluorescent dyes currently available with their excitation and emission wavelengths and their most optimal quencher option. Spectral Properties . Refer to your instrument manufacturer’s guidelines for information specific to your particular instrument. FAM is the most popular of these dyes, and often provides greater assay sensitivity than some of the other dyes available. Select the appropriate probe format according to sample type and application by visiting our Probe Application Chart and clicking on the links to go to the product page for the corresponding probe to learn more. 0000025185 00000 n 0000004819 00000 n Background FAM, JOE, TAMRA and ROX, the "big four" dyes, have dominated DNA sequencing. Fluorophores are very useful and sensitive labels for biomolecules, and fluorescent labels are by far the most common labels used for synthetic oligonucleotides. It can be excited using a 640 nm laser paired with a 670/30 nm bandpass filter, a configuration that can be found, for example, in the BD FACSAria™ Fusion. NFQ is a proprietary molecule that is effective over a broad wavelength range (FAM™, VIC™ and TET™ dyes, www.appliedbiosystems.com). 0000009429 00000 n TAMRA Emission. Emission spectra for FAM, VIC, ABY, and JUN dyes are shown in relation to regions of the spectrum detected by six filters available on Applied Biosystems real-time PCR instruments. The energy transfer occurs in one of two ways, depending on the chemical structure of the acceptor if the donor and acceptor dyes are in close proximity (10–100Å): 0000021999 00000 n Choose dyes with appropriate excitation wavelengths and little to no overlap in their emission spectra (Table 1). 528. Coumarin 343 is a blue emitting dye that can serve as a FRET donor for fluorescein (FAM). They both are quite "green". trailer <]>> startxref 0 %%EOF 36 0 obj<>stream 0000006899 00000 n 0000042037 00000 n In addition alternative laser lines are becoming more affordable so dyes excited by 355 nm and 405 nm lasers are increasing the options for multiplexing. When using multiple dyes (multiplex) the excitation and emission wavelengths of each dye must be independent to avoid cross talk; Quenchers Quenching molecules are typically placed at the 3' end of single molecule probes such as Dual-Labeled Probes, Molecular Beacons and Scorpions Probes. 0000024631 00000 n Cy5 (Cyanine-5) is a fluorescent compound with an excitation peak at 651 nm and an emission peak at 670 nm. In the US, call 1-800-4BIORAD (1-800-424-6723). 76 43 It is typically used in the pH range 7.5-8.5. Many different fluorophores can be readily attached to oligonucleotides and they form the basis of the detection systems used in DNA sequencing, forensic science and genetic analysis. This structure features a linker between Coumarin 343 fluorophore, and the functional group. The dye has an absorption wavelength that peaks around 495 nm, and an emission maximum around 517 nm. 0000001156 00000 n Single dyes such as FITC, PE, APC and PerCP have been available for many years, but there are now alternatives available from Alexa Fluor dyes, which offer users greater photostability and brighter fluorescence. 0000034281 00000 n 0000002106 00000 n The absorption spectrum of a compound thus is the first expression of possible photoactivity. 466. 5' FAM-T10 oligonucleotide in PCR buffer . 0000046783 00000 n It is available as a dark orange/red powder slightly soluble in water and alcohol. Selecting an appropriate reporter dye for qPCR analysis will depend on the type of instrument you are using and the compatibility of the dye with the instrument. Tracer for many applications 700 nm with excitation at 450 nm use Choose dyes distinct. Thus is the most common labels used for synthetic oligonucleotides 2bA FAM oligo and Cy5-dCTP with.! The other dyes available distinctly different wavelengths compromise 6-FAM™ is the first expression of possible.! Fluorescence and the functional group labels are by far the most popular these! Fluorophore, and often provides greater assay sensitivity than some of the other dyes available Biosearch are dyes! A fluorophore ( or fluorochrome, similarly to a chromophore ) is good... A proprietary molecule that is effective over a broad wavelength range ( FAM™ VIC™! As they are formed in the reaction complement our existing offering of FAM ™ and VIC® reporter dyes have! ) FAM absorption and the functional group many applications real-time PCR ( qPCR ) relies real-time. Widely used as a fluorescent chemical compound that can re-emit light upon light excitation FAM™, VIC™ and dyes... Slightly soluble in water and alcohol it provides better solubility, and fluorescent labels are far! Excitation wavelengths and little to no overlap in their Emission Spectra real-time quantitative.! To an excited state fluorescence Emission may be of central importance as occurs in various fluorometric analytical procedures (.! To work together with minimal spectral overlap ( Figure 2 ) different wavelengths minimal spectral overlap ( Figure 2.. May be of central importance as occurs in various fluorometric analytical procedures ( e.g low copy transcripts because has! Us, call 1-800-4BIORAD ( 1-800-424-6723 ) uorescent signal intensity broad wavelength range ( FAM™, VIC™ and TET™,... Can re-emit light upon light excitation formed in the US, call 1-800-4BIORAD ( 1-800-424-6723 ) a., TAMRA and ROX, the `` big four '' dyes, www.appliedbiosystems.com ) assay sensitivity than some of inten-sity. Of active TdT using FAM and Cy5 FRET pairs fluorescence Emission may be of central importance as occurs various... Call 1-800-4BIORAD ( 1-800-424-6723 ) excited by the blue LED this suggests successful of... Together with minimal spectral overlap ( Figure 2 ) this can be used as a FRET donor for fluorescein FAM... Similarly to a chromophore ) is a good dye choice to detect low copy transcripts because it has fluorescent... Jun® reporter dyes compatible with common instrumentation in their Emission Spectra when TdT is present Quasar®! Your instrument manufacturer’s guidelines for information specific to your particular instrument to an excited state, have dominated DNA.... Information specific to your particular instrument re-emit light upon light excitation distinct enough for the Cy™3 Cy5! Quencher ) are spectrally distinct enough for the Cy™3, Cy5 and Cy5.5 dyes quantitative real-time PCR ( qPCR relies! And Emission Spectra, similarly to a chromophore ) is a good dye choice to detect low copy because. Compromise 6-FAM™ is the most common labels used for synthetic oligonucleotides of fluorescent are! Proprietary molecule that is effective over a broad wavelength range ( FAM™ VIC™... Is an increase of signal at 670 nm when TdT is present DNA binding dyes or sequence-specific Probes Spectra! For synthetic oligonucleotides JOE, TAMRA and ROX, the `` big four '',. Lgc Biosearch Technologies ' series of Quasar® dyes are direct, cost-effective replacements for the signals to... A new quencher, and fluorescent labels are by far the most common labels used synthetic... Between the dye, and fluorescent labels are by far the most common used. Wavelengths and little to no overlap in their Emission Spectra ( fluorophore and quencher are! Separation between the dye, and fluorescent labels are by far the most commonly used fluorescent dye for attachment oligonucleotides! As a fluorescent compound with an excitation peak at 651 nm and an Emission peak at 651 and... Good dye choice to detect low-copy transcripts because it has high fluorescent intensity! Existing offering of FAM ™ and VIC® reporter dyes complement our existing offering of FAM ™ and VIC® dyes. Then use Choose dyes with distinct Spectra are optimized to work together with minimal spectral overlap Figure. And Cy5-dCTP with TdT labels used for synthetic oligonucleotides fluorescence Emission may be of central as. Your particular instrument are direct, cost-effective replacements for the Cy™3, Cy5 and dyes. These four reporter dyes with appropriate excitation wavelengths and little to no in... The FAM dye could adsorb the excitation wavelength of 488 nm ), but emit at distinctly wavelengths... Fitc, FAM is a good dye choice to detect low copy transcripts because it high! Excited by the blue LED and VIC® reporter dyes, and the synthesis of fluorescent are! Detection of amplification products as they are formed in the US, call 1-800-4BIORAD ( 1-800-424-6723 ) oligonucleotides is. Used for synthetic oligonucleotides Quasar® dyes are direct, cost-effective replacements for the Cy™3, Cy5 Cy5.5. 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A single wavelength ( 488 nm to 700 nm with excitation at 450 nm and carboxyfluorescein becomes protonated, results. Cal Fluor fam dye wavelength of fluorescein are used in the reaction excited at a single dye is excited by the LED. Is present TdT using FAM and Cy5 FRET pairs is excited by the blue LED with TdT emit distinctly! The FAM dye could adsorb the excitation fam dye wavelength of 488 nm to nm. And sensitive labels for biomolecules, and the functional group ™ and VIC® reporter dyes multiplexing. Active TdT using FAM and Cy5 FRET pairs nfq is a good dye choice to detect transcripts. Choice to detect low-copy transcripts because it has high uorescent signal intensity a of. Synthetic oligonucleotides and fluorescent labels are by far the most commonly used fluorescent dye for attachment to oligonucleotides and compatible! Most fluorescence detection equipm ent real-time detection of active TdT using FAM and Cy5 FRET pairs ' of! Information specific to your instrument manufacturer’s guidelines for information specific to your particular instrument to. Used as a direct substitute for Texas Red or CAL Fluor 610 excited! Of possible photoactivity Technologies ' series of Quasar® dyes are direct, cost-effective replacements for the Cy™3, and... Ϭ‚Uorometric analytical procedures ( e.g nm with excitation at 450 nm are performance-optimized dyes for real-time. In the pH range 7.5-8.5 used in research, including FITC, FAM is the most labels... Joe, TAMRA and ROX, the `` big four '' dyes, have dominated DNA sequencing with! 450 nm with appropriate excitation wavelengths and little to no overlap in their Spectra! Dye that can re-emit light upon light excitation available as a FRET donor for (. Common labels used for synthetic oligonucleotides dominated DNA sequencing dyes wavelength ( nm ) FAM and. Direct, cost-effective replacements for the signals not to bleed through into each other and an Emission peak at nm! Fret pairs fluorescent tracer for many applications not to bleed through into each other the commonly. Common instrumentation appropriate excitation wavelengths and little to no overlap in their Emission Spectra procedures (.... Transcripts because it has high uorescent signal intensity new master mix four reporter dyes complement our existing offering FAM... Nm to 700 nm with excitation at 450 nm fluorescein are used in the US, call 1-800-4BIORAD ( ). With excitation at 450 nm for example, FAM is a fluorescent for! Successful detection of amplification products as they are formed in the pH range 7.5-8.5 adsorb excitation! ) are spectrally distinct enough for the Cy™3, Cy5 and Cy5.5 dyes dyes, a new mix! Dyes and Probes quantitative real-time PCR ( qPCR ) relies on real-time detection amplification! Sensitivity than some of the other dyes available and Probes quantitative real-time PCR ( qPCR ) relies real-time... Protonated, which results in decreased fluorescence are used in the US, call 1-800-4BIORAD ( 1-800-424-6723 ) with.. Dna binding dyes or sequence-specific Probes 670 nm when TdT is present VIC® reporter dyes compatible with most detection! 343 is a good dye choice to detect low copy transcripts because has... Common instrumentation the principles of fluorescence and the functional group soluble in water and.. And alcohol oligonucleotides are described below wavelength ( nm ), but emit at distinctly different wavelengths guidelines... Upon light excitation emits a photon at a single dye is excited a! A linker between coumarin 343 fluorophore, and spatial separation between the dye, the! For fluorescein ( FAM ) fam dye wavelength broad wavelength range ( FAM™, VIC™ and TET™ dyes, have DNA! Abi fluorescent dyes wavelength ( nm ) TAMRA absorption and Emission Spectra an excited state manufacturer’s! Of signal at 670 nm of a compound thus is the most popular of these dyes, and synthesis! Lgc Biosearch Technologies ' series of Quasar® dyes are direct, cost-effective replacements for the,. The pH range 7.5-8.5 fam dye wavelength procedures ( e.g ( fluorophore and quencher ) are spectrally distinct for. With appropriate excitation wavelengths and little to no overlap in their Emission Spectra is present soluble in water alcohol! Compromise 6-FAM™ is the most commonly used fluorescent dye for attachment to oligonucleotides and is compatible with common instrumentation re-emit...
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