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Oligo Synthesis

Oligo Synthesis : Reagents, Labelling

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Expedite 8909 and Cyclone (240mL capacity)

Expedite 8909 and Cyclone (240mL capacity)

Glen Research

Description

Expedite 8909 and Cyclone (240mL capacity)

Structure

Catalog Number: 40-4036-xx

Description: Sulfurizing Reagent

3H-1,2-Benzodithiole-3-one-1,1-dioxide
Formula: C7H4O3S2 M.W.: 200.23

Diluent: Anhydrous Acetonitrile

Technical Bulletin
Storage: Freezer storage, -10 to -30°C, dry
Stability in Solution: May precipitate if synthesizer line to bottle is contaminated, otherwise ~1month

SULFURIZING REAGENT

Glen Research’s Sulfurizing Reagents are used to prepare phosphorothioate linkages using CE phosphoramidite chemistry. Each reagent exhibits the following attributes:
1) Reliably soluble, making them safe to use on automated synthesizers.
2) Reaction is fast (30 seconds), making the process convenient on small scales and readily amenable to scale-up.
3) Process is efficient, with better than 96% of the linkages being phosphorothioate and the remainder being phosphodiester.
When ordering Sulfurizing Reagent (Beaucage Reagent), please also order the silanized bottle appropriate for your instrument. There will be no charge for one bottle when supplied with a reagent order. The silanized bottle can be reused after rinsing with acetonitrile and drying. Additional silanized bottles are priced as shown below. Dissolve the reagent in acetonitrile at a concentration of 1g/100mL.

Sulfurizing Reagent II (3-((Dimethylamino-methylidene)amino)-3H-1,2,4-dithiazole-3-thione, DDTT) exhibits all the properties of Beaucage Reagent while adding stability in solution on the synthesizer AND offering strong ability to sulfurize RNA linkages. Sulfurizing Reagent II is available in powder form and as a stable solution.

 

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Protocols

Material Safety Data Sheet

Glen Report 6.2: RESEARCH REVIEW - ANTISENSE RNA

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References

FREQUENTLY ASKED TECHNICAL QUESTION

QUESTION: You say that the sulfurizing reagent, 40-4036, is stable for 1 month in solution "under ideal conditions". What are ideal conditions?

RESPONSE:The sulfurizing reagent, developed by Beaucage, is a very reactive product. In solution in acetonitrile, it must be stored in plastic or silanized glass containers. Solutions in sealed bottles are stable for periods of months, turning cloudy and finally precipitating as decomposition occurs.

On most synthesizers, the sulfurizing reagent must be installed on the port previously used for the iodine oxidizing solution. In this situation, the reagent will decompose quite rapidly over 24 to 48 hours. Indeed, we recommend weighing out the required amount of dry reagent into the silanized bottle provided. The recommended concentration for use is 1g/100mL So, if 40mL of solution is desired, weigh 0.4g of reagent into the silanized bottle and add 40mL of acetonitrile. It is important to flush the synthesizer line liberally first with acetonitrile then with the sulfurizing solution to remove traces of the previous solution from the line. After synthesis, we recommend removing the solution from the instrument immediately and discarding the remaining amount.

On synthesizers which have a spare port to accommodate an additional liquid reagent, the stability of the sulfurizing solution will progressively improve until the solution can be left on the machine for at least 1 month.

REFERENCE(S):
R.P. Iyer, W. Egan, J.B. Regan, and S.L. Beaucage, J. Amer. Chem. Soc., 1990, 112, 1253-1254.


QUESTION: Why can't I see my phosphorothioate oligo when I stain the gel with ethidium bromide?

RESPONSE:It appears that the sulfur of the phosphorothioate linkage can quench the fluorescence of ethidium bromide. I know researchers who could see their phosphorothioate oligo by UV shadowing but when stained with EtBr, no bands were observed, while on the same gel, normal phosphodiester oligos stained perfectly.

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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
0.05M Dil.
(mL)
LV40 LV200 40nm 0.2µm 1µm 10µm
Approximate Number of Additions
40-4036-10 1gram 1grams N/A N/A N/A N/A N/A N/A N/A
40-4036-20 2grams 2grams N/A N/A N/A N/A N/A N/A N/A
Expedite
Cat.No. Pack
Size
Grams/
Pack
Dilution
(mL)
Molarity 50nm 0.2µm 1µm 15µm
Approximate Number of Additions
40-4036-10 1gram 1grams N/A N/A N/A N/A N/A
40-4036-20 2grams 2grams N/A N/A N/A N/A N/A
Beckman
Cat.No. Pack
Size
Grams/
Pack
Dilution
(mL)
Molarity 30nm 200nm 1000nm

Approximate Number of Additions
40-4036-10 1gram 1grams N/A N/A N/A N/A
40-4036-20 2grams 2grams N/A N/A N/A N/A

If you cannot find the answer to your problem then please contact us or telephone +44 (0)1954 210 200

Related products

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