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AAT Bioquest

Custom Oligonucleotide Labeling

AAT Bioquest offers a wide range of modifications for DNA and RNA oligonucleotides, including fluorescent reporters, quencher labels, hapten modifications, as well as, amine and thiol functional linkers. Our custom services are designed to meet your specifications with stringent quality control and quick turnaround, to supply scientists with the necessary tools to satisfy even the most challenging of applications. Several applications in which our molecular biology tools can be successfully employed include fluorescence in situ hybridization (FISH), blotting assays, cell trafficking, intracellular localization, affinity purification, PCR, microarrays and sequencing.

 

Guaranteed quality

Affordable price tiers that work with your budget

Scalable service with minimum order of 250 nmole

Quick order fulfillment modified oligonucleotides delivered in 3-5 business days*


Oligonucleotides for modification must be supplied by the inquirer, AAT Bioquest does not custom manufacture any oligonucleotides. Contact us for a quote today, and let our highly motivated and experienced team of scientists work for you! Oligonucleotide Modifications

Modified oligonucleotides are utilized in a wide range of applications spanning the fields of biotechnology, molecular biology, diagnosis and drug discovery. Depending upon the coupled moiety or reporter molecule, modified oligonucleotides can be used to provide a particular reactivity linking an oligonucleotide either to another molecule or desired surface, or can be modified to enhance visualizaton. Common modifications include biotin, digoxigenin, dinitriphenyl, amino-modifiers, alkyne modifiers, phosphate modifiers, fluorescent reporters and quencher labels.

Biotin-modified oligonucleotides, in particular, bind tightly and with high affinity to streptavidin. The streptavidin can be conjugated to a reporter molecule, such as a fluorophore or enzyme, and together they can been used in a variety of biological assays and affinity purification applications. Biotinylated oligonucleotides can also be used as probes or primers in either in vivo or in vitro applications. In PCR, the addition of 3'-phosphate groups block elongation, while in functional assays the incorporation of reactive moieties facilitate the covalent linking of nonradioactive labels such as enzymes. Several enzymes, such as horseradish peroxidase, are compatible with standard hybridization conditions, and allow detection by a number of endpoints including absorbance, fluorescence and luminescence.

Oligonucleotides can also be fluorescently modified. Oligos bearing fluorescent tags have been widely used as probes in automated DNA sequencing, FISH and PCR product detection. Dual-modified oligonucleotides with a fluorophore and a quencher at their 5' and 3' ends, respectively, have been used in all major types of fluorescence imaging including fluorescence resonance energy transfer (FRET) and quantitative PCR. In the presence of a complementary sequence, these probes commonly referred to as molecular beacons, unfold and hybridize to the target. The resulting conformational change produces sufficient displacement of the fluorophore from its respective quencher, thereby restoring fluorescence. In quantitative PCR, molecular beacons bind to the amplified target following each cycle of amplification and the resulting signal is proportional to the amount of template. Common fluorophore/quencher pairs include EDANS/DABCYL and Tide Fluor™-Tide Quencher™ pairs.

Available Labels

Picking the correct label can be challenging. Let our experts help simplify the process! Contact us by email or phone (1-(800)-990-8053) for a consultation and let us work with you to pick the best label to suit your specific needs. And don't fret if you can't find a modification you need on our website, AAT Bioquest offers several modifications that are not listed in our online catalog.

TypeLabelApplications
EnzymeHorseradish Peroxidase (HRP)HRP-oligos requires substrates for detection, such as TMB or Amplite® Red. It can be used in in situ hybridization, dot blots, cellular localization and non-radioactive immunoassays.
Alkaline Phosphatase (AP)AP-oligos require substrates for detection, such as FDP or MUP. It can be used in in situ hybridization, dot blots, cellular localization and non-radioactive immunoassays.
Other TagsBiotinBiotin-streptavidin detection system can be used in oligonucleotide labeling, in situ hybridization, blot hybridization, matrix binding, cellular localization, genomic hybridization, affinity binding and purification.
Digoxigenin (DIG)DIG binds tightly and with high affinity to anti-DNP antibodies. It can be used in oligonucleotide labeling, in situ hybridization, blot hybridization, cellular localization, genomic hybridization, affinity binding and purification.
Dinitriphenyl (DNP)DNP binds tightly and with high affinity to anti-DNP antibodies. It can be used in oligonucleotide labeling, in situ hybridization, blot hybridization, cellular localization, genomic hybridization, affinity binding and purification.
3' and 5' ModificationAmine-ModifiersAddition of primary amino groups (-NH?) facilitates the attachment of various modifiers including non-fluorescent tags, enzymes, fluorescent dyes and quenchers. It can also be used to attach an oligonucleotide to a solid surface.
Thiol-ModifiersAddition of thiol groups (-SH) facilitates the attachment of various modifiers including non-fluorescent tags, enzymes, fluorescent dyes and quenchers. It can also be used to attach an oligonucleotide to a solid surface.
Alkyne-ModifiersAddition of alkyne groups facilitates the attachment of various fluorescent and non-fluorescent modifiers via click-chemistry.
Phosphate-ModifiersAddition of 3'-phosphate groups inhibits degradation by some nucleases or blocks elongation by DNA polymerases. Addition of 5'-phosphate groups to oligos allows for subsequent ligation by DNA ligases.
 

Table 1. Fluorescent Labels for Oligo Labeling

Emission Color
Conjugate
Ex (nm)
Em (nm)
ε¹
CF 260 nm²
 AMCA35345519,0000.183
 EDANS3364555,9000.34
 6-FAM49452175,0000.255
 6-TET52153676,0000.191
 6-JOE52054873,0000.326
 6-HEX53355074,0000.300
 Cy3549565150,0000.033
 Cy3.5581569125,0000.151
 6-TAMRA54757375,0000.335
 6-TAMRAA CPG *1000 Å*54757375,0000.335
 5(6)-TAMRA54657578,0000.320
 5(6)-ROX56859193,0000.301
 Texas Red®58860195,0000.456
 6-ROX57560295,0000.307
 Cy5644665250,0000.088
 Cy5.5678701230,0000.094
 Cy7749776275,0000.025
 ICG784813230,0000.112
  1. ε = Extinction coefficient (cm-1M-1) at their maximum absorption wavelength
  2. CF at 260 nm is the correction factor used for eliminating the dye contribution to the absorbance at 260 nm (for oligos and nucleic acid labeling)

Table 2. AAT Bioquest Fluorescent Tide Fluor™ Dyes

Emission Color
Conjugate
Ex (nm)
Em (nm)
ε¹
CF 260 nm²
 Tide Fluor™ 134542220,0000.246
 Tide Fluor™ 2WS50252575,0000.211
 Tide Fluor™ 250052775,0000.288
 Tide Fluor™ 3WS555565150,0000.079
 Tide Fluor™ 354657185,0000.331
 Tide Fluor™ 459061890,0000.489
 Tide Fluor™ 5WS649664250,0000.023
 Tide Fluor™ 6WS676695220,0000.111
 Tide Fluor™ 7WS749775275,0000.009
 Tide Fluor™ 8WS775807250,0000.103
  1. ε = Extinction coefficient (cm-1M-1) at their maximum absorption wavelength
  2. CF at 260 nm is the correction factor used for eliminating the dye contribution to the absorbance at 260 nm (for oligos and nucleic acid labeling)

Table 3. AAT Bioquest Non-Fluorescent Dyes and Quenchers

  1. ε = Extinction coefficient (cm-1M-1) at their maximum absorption wavelength
  2. CF at 260 nm is the correction factor used for eliminating the dye contribution to the absorbance at 260 nm (for oligos and nucleic acid labeling)

Please contact us at info@aatbio.com to inquire about additional or custom labels.


Additional Resources
 

Table 4. Tide Fluor™ equivalents of common dyes

If you are using
Try this Tide Fluor™ dye
EDANSTide Fluor™ 1 (TF1)
FAM, FITC, Alexa Fluor® 488Tide Fluor™ 2 (TF2)
Alexa Fluor® 488Tide Fluor™ 2WS (TF2WS)
Cy3®, Alexa Fluor® 555Tide Fluor™ 3 (TF3)
Cy3®, Alexa Fluor® 555Tide Fluor™ 3WS (TF3WS)
ROX, Texas Red®, Alexa Fluor® 594Tide Fluor™ 4 (TF4)
Cy5®, Alexa Fluor® 647Tide Fluor™ 5WS (TF5WS)
Cy5.5®, IRDye® 700, Alexa Fluor® 680Tide Fluor™ 6WS (TF6WS)
Cy7®, Alexa Fluor® 750Tide Fluor™ 7WS (TF7WS)
IRDye® 800Tide Fluor™ 8WS (TF8WS)

Table 5. Tide Quencher™ equivalents of common dyes


Table 6. Recommended FRET pairs for developing FRET oligonucleotides

Donor \ Acceptor
DABCYL
TQ1
TQ2
TQ3
TQ4
TQ5
TQ6
TQ7
EDANS+++++++-----
MCA+++++++-----
Tide Fluor™ 1+++++++-----
FAM
FITC
++++++----
Cy2®
Tide Fluor™ 2
++++++----
HEX
JOE
TET
--+++++---
Cy3®
TAMRA
Tide Fluor™ 3
--+++++---
ROX
Texas Red®
---+++++--
Tide Fluor™ 4---+++++--
Cy5®
Tide Fluor™ 5
----+++++-
Cy5.5®
Tide Fluor™ 6
-----+++++
Cy7®
Tide Fluor™ 7
------++++
Optimal choice
Acceptable to use
Not recommended to use