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Oligo Synthesis

Oligo Synthesis : CEPs

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N4-Et-dC-CE Phosphoramidite

N4-Et-dC-CE Phosphoramidite

Glen Research

Catalogue No.DescriptionPack SizePriceQty
10-1068-02N4-Et-dC-CE Phosphoramidite 0.25g £673.00 Quantity Add to Order
10-1068-90N4-Et-dC-CE Phosphoramidite 100µmoles £227.00 Quantity Add to Order
10-1068-95N4-Et-dC-CE Phosphoramidite 50µmoles £127.00 Quantity Add to Order

Description

N4-Et-dC-CE Phosphoramidite

Structure

Catalog Number: 10-1068-xx

Description: N4-Et-dC-CE Phosphoramidite

5'-Dimethoxytrityl-N4-ethyl-2'-deoxyCytidine,3'-[(2-cyanoethyl)-
(N,N-diisopropyl)]-phosphoramidite
Formula: C41H52N5O7P M.W.: 757.87 F.W.: 317.42

Diluent: Anhydrous Acetonitrile
Coupling: No changes needed from standard method recommended by synthesizer manufacturer.
Deprotection: No changes needed from standard method recommended by synthesizer manufacturer.
Storage: Refrigerated storage, maximum of 2-8°C, dry
Stability in Solution: 2-3 days

BASES AFFECTING DUPLEX STABILITY

C-5 methyl pyrimidine nucleosides are known to stabilize duplexes relative to the non-methylated bases. Therefore, enhanced binding can be achieved using 5-methyl-dC in place of dC, duplex melting temperature being increased by 1.3°. Improved stacking in this case is believed to be brought about by elimination of water molecules from the duplex. 2,6-Diaminopurine 2'-deoxyriboside (2-amino-dA) forms an additional hydrogen bond with Thymidine, thereby leading to duplex stabilization with a melting temperature increase of 3°. Our 2-amino-dA monomer exhibits fast and effective deprotection in ammonium hydroxide and it is stabilized to depurination during synthesis. Sequences with high GC content may contain mismatches and still hybridize because of the high stability of the G-C base pair. The N4-ethyl analogue of dC (N4-Et-dC) hybridizes specifically to natural dG but the stability of the base pair is reduced to about the level of an AT base pair.

AP-dC (G-clamp) enhances oligo hybridization since the AP-C....G base pair contains 4 hydrogen bonds, which makes the interaction much stronger than the regular C....G base pair with its 3 hydrogen bonds.

Sequences with high GC content may contain mismatches and still hybridize because of the high stability of the G-C base pair. The N4-ethyl analogue of dC (N4-Et-dC) hybridizes specifically to natural dG but the stability of the base pair is reduced to about the level of an AT base pair.

Coupling N6-Me-dA (10-1003) and N4-Et-dC (10-1068) with 1H-tetrazole leads to a trace of branching at the secondary amine positions, while DCI leads to around 15% branching. In collaboration with Berry and Associates, the acetyl protected monomers were prepared. Acetyl protection was chosen since it would block branching reactions. Oligonucleotides synthesized using these monomers proved to be compatible with all popular deprotection strategies from UltraMild to UltraFast. When the acetyl protected monomers were compared with the unprotected monomers using DCI as activator, branching was reduced from 15% to zero.

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Protocols

Material Safety Data Sheet

??Glen Report 11.1: USING MODIFIED BASES TO OPTIMIZE HYBRIDIZATION

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Applications & Benefits

DILUTION/COUPLING DATA

The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures. Please link for more detailed usage information with the various synthesizers.

ABI 392/394
Cat.No. Pack
Size
Grams/
Pack
0.1M Dil.
(mL)
LV40 LV200 40nm 0.2µm 1µm 10µm
Approximate Number of Additions
10-1068-02 0.25grams .25grams 3.3 96.67 58 36.25 26.36 19.33 4.83
10-1068-90 100µmoles .076grams 1 20 12 7.5 5.45 4 1
10-1068-95 50µmoles .038grams .5 3.33 2 1.25 .91 .67 .17
Expedite
Cat.No. Pack
Size
Grams/
Pack
Dilution
(mL)
Molarity 50nm 0.2µm 1µm 15µm
Approximate Number of Additions
10-1068-02 0.25grams .25grams 4.92 .07 92 57.5 41.82 5.75
10-1068-90 100µmoles .076grams 1.5 .07 23.6 14.75 10.73 1.48
10-1068-95 50µmoles .038grams .75 .07 8.6 5.38 3.91 .54
Beckman
Cat.No. Pack
Size
Grams/
Pack
Dilution
(mL)
Molarity 30nm 200nm 1000nm

Approximate Number of Additions
10-1068-02 0.25grams .25grams 4.92 .07 93.6 58.5 42.55

10-1068-90 100µmoles .076grams 1.5 .07 25.2 15.75 11.45

10-1068-95 50µmoles .038grams .75 .07 10.2 6.38 4.64

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Related products

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