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Oligo Modifications List | Oligo Modifications Reference Category
Modification : Digoxigenin NHS
Reference Catalog Number 26-6429
Category Affinity Ligands
Modification Code Dig-N
5 Prime Y
3 Prime Y
Internal Y
Molecular Weight (mw) 561.3
Extinction Coeficient (ec) 13
Technical Info (pdf) PS26-6429.pdf
Catalog NoScalePrice
26-6429-0550 nmol$465.00
26-6429-02200 nmol$465.00
26-6429-011 umol$987.00
26-6429-032 umol$1,536.00
26-6429-1010 umol$4,584.48
26-6429-1515 umol$6,355.60
Related Modifications
Amino C6
Amino dA C6
Amino Serinol
BiotinTEG Azide
PC Biotin (photocleavable)
Amino dT C6
Amino dG C6
Amino C3
Biotin NHS
Amino C12
Biotin dT
Biotin Multi
DNP TEG (2, 4-dinitrophenyl)
Biotin TEG
Amino dC C6

This modification is a post synthesis conjugation to a primary amino group thus an additional modification with an amino group is required. A C3, C6 or C12 amino group can be placed at the 5' or for the 3' end a C3 or C7 amino and for internal positions an amino modified base is used, e.g Amino dT C6.

NHS based modifications are post synthesis conjugation performed using a primary amino group. The yield is lower as compared to direct automated coupling of modifications that are available as amidites. Approximate yield for various scales are given below.

~2 nmol final yield for 50 nmol scale synthesis.
~5 nmol final yield for 200 nmol scale synthesis.
~16 nmol final yield for 1 umol scale synthesis

Digoxigenin (as Digoxigenin-3-O-methylcarbonyl-epsilon-aminocaproic acid NHS ester) is a member of the steroid family found in Digitalis plants (1). It is a hapten, that is, a small molecule having high immunogenicity. Because antibodies raised against haptens have considerably higher affinities for them than other antibodies do for their targets makes haptens particularly desirable as affinity tags for oligonucleotides (2).

Digoxigenin ('Dig') is commonly used to label oligonucleotides probes for use in hybridization applications, for example, in situ hybridization (3), Northern and Southern blotting. After hybridization to their targets, these Dig-labeled probes are detected with anti-Dig antibodies that are labeled with dyes (for primary detection) or enzymes (for secondary detection using a fluorogenic, chemiluminogenic, or colorimetric substrate specific for the enzyme). To maximize signal, Gene Link recommends modifying the oligonucleotide probe with three Dig molecules, spaced about 10 bases apart. Note that since digoxigenin is in the form of an NHS ester, an active primary amino group (such as Amino Linker C6) must first be incorporated into the oligonucleotide, to allow for subsequent conjugation to the digoxigenin NHS ester.

1. Polya, G. Biochemical targets of plant bioactive compounds. New York: CRC Press, 2003. p 847.
2. Shreder, K. Synthetic Haptens as Probes of Antibody Response and Immunorecognition. Methods (Academic Press) (2000), 20: 372-379.
3. Hauptmann, G., Gerster, T.. Two-color whole-mount in situ hybridixation to vertebrate and Drosophila embryos.Trends Genet. (1994), 10: 266.

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