Asymmetric Doubler (Lev) Phosphoramidite - (10-1981)
1-[5-(4,4’-dimethoxytrityloxy)pentylamido]-3-[5-levulinyloxypentylamido]-propyl-2-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
1-[5-(4,4’-dimethoxytrityloxy)pentylamido]-3-[5-levulinyloxypentylamido]-propyl-2-[(2-cyanoethyl)-(N,N-diisopropyl)]-phosphoramidite
Dendrimers are discrete, highly branched, monodispersed polymers that possess patterns reminiscent of the branching of trees. Plain and mixed oligonucleotide dendrimers can be synthesized using novel doubling and trebling phosphoramidite synthons.1,2 Dendrimers offer the following advantages. Incorporation of label using γ-32P-ATP and polynucleotide kinase increases in proportion to the number of 5’-ends. Fluorescent signal also increases in proportion to the number of 5’-ends, if spacers are incorporated between the labels and the ends of the branches. When using a dendrimeric oligonucleotide as a PCR primer, the strand bearing the dendrimer is resistant to degradation by T7 Gene 6 exonuclease making it easy to convert the double-stranded product of the PCR to a multiply labeled, single-stranded probe. Enhanced stability of DNA dendrimers makes them useful as building blocks for the ‘bottom up’ approach to nano-assembly. These features also suggest applications in DNA chip technology when higher temperatures are required, for example, to melt secondary structure in the target.
Usage
- Coupling: 6 minute coupling time recommended.
- Deprotection: Levulinyl The levulinyl protecting group can be selectively removed without cleavage of the oligonucleotide from the CPG by treatment with 0.5M Hydrazine hydrate in 1:1 pyridine/acetic acid. [Note hydrazine hydrate is a violent poison that is both volatile and readily absorbed through skin] Fit the column with syringes and push the solution back and forth across the column. Let sit for 15 minutes at room temperature. Rinse the column with 1.5 mL of 1:1 pyridine/acetic acid (3x) and then 1.5 mL of ACN (3x). Dry CPG under a stream of argon and proceed with the synthesis of the branching sequence. If a non-branching control is desired, simply deprotect in ammonium hydroxide as required by the nucleobases.
Specifications | |
---|---|
Diluent | Anhydrous Acetonitrile |
Recommended Storage | Freezer storage, -10 to -30°C, dry |
Stability In Solution | 1-2 Days |
The table below show pack size data and, for solutions, dilution and approximate coupling based on normal priming procedures.
ABI 392/394
Catalog # | Pack Size | Grams/Pack | 0.1M Dil. (mL) | Approximate Number of Additions | |||||
---|---|---|---|---|---|---|---|---|---|
LV40 | LV200 | 40nm | 0.2μm | 1μm | 10μm | ||||
10-1981-02 | 0.25 g | 0.25 | 2.81 | 80.33 | 48.2 | 30.13 | 21.91 | 16.07 | 4.02 |
10-1981-90 | 100 µmol | 0.09 | 1 | 20 | 12 | 7.5 | 5.45 | 4 | 1 |
Expedite
Catalog # | Pack Size | Grams/Pack | Dilution (mL) | Approximate Number of Additions | ||||
---|---|---|---|---|---|---|---|---|
Molarity | 50nm | 0.2μm | 1μm | 15μm | ||||
10-1981-02 | 0.25 g | 0.25 | 4.19 | 0.07 | 77.4 | 48.38 | 35.18 | 4.84 |
10-1981-90 | 100 µmol | 0.09 | 1.5 | 0.07 | 23.6 | 14.75 | 10.73 | 1.48 |
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