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Liquid Reagents

In solid-phase synthesis of oligonucleotides, liquid reagents are used in each step of the synthesis cycle. Overall synthesis performance, and therefore total product yield and purity of the crude oligonucleotide, is highly dependent on the chemical purity of the monomers and the supporting liquid reagents.

Proligo Reagents™ has more than 15 years of manufacturing experience in DNA and RNA synthesis reagents. Our liquid reagents meet the most stringent quality specifications, making them ideal for use in automated nucleic acid synthesis. Lot-to-lot consistency is guaranteed for every product. Each lot is individually quality-controlled in our ISO 9001-certified site to ensure optimum synthesis performance.

Chemical Composition and Purpose of Proligo Reagents' Liquid Reagents During the Synthesis Cycle

Deblock solution,  

containing trichloroacetic acid (TCA) or dichloroacetic acid (DCA) in dichloromethane, removes the dimethoxytrityl (DMT) protecting group from the 5' hydroxyl moiety of nucleotides already incorporated into the growing nucleic acid, prior to the addition of the next phosphoramidite. Removal of the DMT allows the unprotected 5' hydroxyl moiety to react with a new phosphoramidite in a subsequent extension reaction.

Wash solution, 

containing anhydrous acetonitrile (water content < 30ppm), is used as the overall wash solvent following each step in the synthesis cycle.

Activator solution,

containing 4,5-dicyanoimidazole (DCI) in acetonitrile, is typically added simultaneously with a phosphoramidite during the extension step. The activator reacts with the trivalent phosphorous of the phosphoramidite to form a reactive intermediate. This intermediate then forms an internucleotide phosphite with the detritylated 5' hydroxyl group of the growing oligonucleotide chain.

Activator 42™

Cap A solution, 

containing acetic anhydride and tetrahydrofuran (THF), is used after the phosphoramidite reaction/coupling step. This solution aborts chains bearing unreacted 5' hydroxyl groups due to failure of reaction with activated phosphoramidite (typically, 1 to 2% of growing oligonucleotide chains in each synthesis cycle). These unreacted 5' hydroxyl groups are capped with an acetyl group and thus rendered unreactive for subsequent synthesis steps. For synthesis with Proligo Reagents' TAC-protected phosphoramidites, Fast Deprotection CAP A solution must be employed instead of Cap A solution.

Fast Deprotection Cap A solution,

must be used for synthesis with Proligo Reagents' TAC-protected phosphoramidites. Fast Deprotection Cap A solution, containing tert-butylphenoxyacetyl acetic anhydride (tac₂ O) in tetrahydrofuran, is used in place of Cap A solution to ensure that the displacement of tert-butylphenoxyacetyl (TAC) does not occur.

Cap B solution,

containing tetrahydrofuran (THF), pyridine and 1-methylimidazole, is mixed in-situ with Cap A solution during the capping (acetylation) reaction. The pyridine maintains a pH basic enough to promote the acetylation reaction and the 1-methylimidazole, together with the acetic anhydride, makes an excellent acetylating agent.

Oxidizer solution,

containing tetrahydrofuran (THF), pyridine, iodine and water, is used to oxidize the chemically unstable trivalent phosphorous triester linkage after the coupling reaction to the stable pentavalent phosphate triester linkage of natural nucleic acids. Here, iodine functions as a mild oxidant and water functions as an oxygen donor.

Product List for Liquid Reagents

Other quantities are available upon request.

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