Glen Research Product Information

Catalog Number: 10-1038-xx
Description: Biotin-dT

5'-Dimethoxytrityloxy-5-[N-((4-t-butylbenzoyl)-biotinyl)-aminohexyl)- 3-acrylimido]-2'-deoxyUridine-3'-[(2-cyanoethyl)-(N,N-diisopropyl)]- phosphoramidite

Formula: C69H89N8O12PS 
M.W.: 1285.55 
F.W.: 684.70 

Diluent: Anhydrous Acetonitrile

Coupling: 15 minute coupling recommended

Deprotection: No changes needed from standard method recommended by synthesizer manufacturer.

Storage: Refrigerated storage, maximum of 2-8°C, dry

Stability in Solution: 3-5 days

   

Pricing Information

Catalog Number

Pack Size

Price (US$)


10-1038-95
50µm
167.50
10-1038-90
100µm
325.00
10-1038-02
0.25g
625.00

Related Document(s)

  

Literature Highlight(s)

FREQUENTLY ASKED TECHNICAL QUESTION

QUESTION:
1. If Biotin-dT is inserted at the 5'-terminus, will it affect the function of kinase?
2. Does Biotin-dT affect hybridization?

RESPONSE:1. Biotin dT will not affect the ability of kinase to phosphorylate at the 5'-terminus.

2. The presence of Biotin-dT within an oligonucleotide does not affect hybridization relative to the same oligo containing dT(1). The biotin resides in the major groove of the formed double-stranded DNA and is readily available for binding to avidin or streptavidin.

REFERENCE(S):
(1) J. Telser, K.A. Cruickshank, L.E. Morrison, and T.L. Netzel, J. Am. Chem. Soc., 1989, 111, 6966-6976.


QUESTION: What absorbance does biotin have at 260nm? The HPLC trace shows absorbance at 254nm?

RESPONSE:Biotin is transparent at 260nm. The UV detector in the HPLC trace of biotin phosphoramidites is monitoring the absorption of the DMT group on the spacer arm.


QUESTION: Do you have a biotin phosphoramidite containing a disulfide linker which can be cleaved later with DTT to release the DNA from a streptavidin support?

RESPONSE:No. However, this can be produced on the synthesizer by adding to the 5'- terminus first 5'-thiol-modifier C6 S-S (10-1936) followed by BioTEG phosphoramidite (10-1955). This should generate a biotinylated primer with a long spacer arm containing the disulfide linkage which can be cleaved later with DTT.


QUESTION: How can I tell if the biotinylated oligonucleotide I have made really does contain biotin?

RESPONSE:A colorimetric assay for biotin can be quite effective. The color results from the reaction of biotin with p-dimethylaminocinnamaldehyde in the presence of sulfuric acid.

1. Spot 0.2 A260 units of biotinylated oligonucleotide on a silica gel TLC plate.

2. Dry the plate.]

3. Spray with a solution of 2% p-dimethylaminocinnamaldehyde (Sigma), 2% conc. sulfuric acid in ethanol.

4. Heat the plate and the presence of biotin will be indicated by the formation of a pink spot.

Since the intensity of the biotin spot is quite low, it is prudent to compare with an unlabelled oligonucleotide similarly treated.


DILUTION/COUPLING DATA

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)
LV40LV20040nm0.2µm1µm10µm
Approximate Number of Additions
10-1038-020.25grams.25grams1.9451.3330.819.251410.272.57
10-1038-90100µmoles.129grams120127.55.4541
10-1038-9550µmoles.064grams.53.3321.25.91.67.17
Expedite
Cat.No.Pack
Size
Grams/
Pack
Dilution
(mL)
Molarity50nm0.2µm1µm15µm
Approximate Number of Additions
10-1038-020.25grams.25grams2.9.0751.632.2523.453.23
10-1038-90100µmoles.129grams1.5.0723.614.7510.731.48
10-1038-9550µmoles.064grams.75.078.65.383.91.54

 

 

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01/18/2019

 

 

 

 

 

 

 

 

 

 

 

 

 

 


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