Abstract: Flow control mechanisms control the direction and flow rate of synthesis reagent through one or more synthesis reaction vessels for automated solid phase synthesis. Selectable, known, and reproducible positive or negative pressure differentials (?5 to +10 psi) accomplish controlled, bidirectional (forward and reverse) flow of synthesis reagents through synthesis media contained within the reaction vessels. Venturi-based vacuum apparatus, valves, electronic pressure regulators and compound digital pressure gauge, can be added to automated solid phase synthesis instruments to provide, control, and monitor known, selectable, reproducible negative and positive pressures to one or both valve sealable and un-sealable ends (inlets and outlets) of the reaction vessel as needed to generate and reverse said pressure differentials between the opposite ends of said synthesis reaction vessels, yielding controlled forward and backward flows of synthesis reagents through the synthesis media.

Abstract: Flow control mechanisms control the direction and flow rate of synthesis reagent through one or more synthesis reaction vessels for automated solid phase synthesis. Selectable, known, and reproducible positive or negative pressure differentials (?5 to +10 psi) accomplish controlled, bidirectional (forward and reverse) flow of synthesis reagents through synthesis media contained within the reaction vessels. Venturi-based vacuum apparatus, valves, electronic pressure regulators and compound digital pressure gauge, can be added to automated solid phase synthesis instruments to provide, control, and monitor known, selectable, reproducible negative and positive pressures to one or both valve sealable and un-sealable ends (inlets and outlets) of the reaction vessel as needed to generate and reverse said pressure differentials between the opposite ends of said synthesis reaction vessels, yielding controlled forward and backward flows of synthesis reagents through the synthesis media.

Abstract: Non-hydrophobic beads and methods to reversibly bind, normalize, store and in situ deliver primers to reactions including PCR. Also provided are instructions for preparing the beads. In the presence of an appropriate binding buffer, a bead can be used to bind and desalt primers from a crude solution of DMT-off primers. In the presence of an appropriate binding buffer, a bead can be used to bind and purify primers from a crude solution of DMT-on primers. A bead may bind a picomolar amount of DMT-on primers from a solution containing a plurality of crude DMT-on primers. Upon detritylation and washing, the resulting DMT-off primer bound bead may be used in PCR. Primers are released from the bead upon cycling the temperature. Primer bound beads are coated or silanized with hydrophobic reagents which ensures a gradual release of primers during the thermal cycling of the PCR reaction. Coating or silanization in turn enhances primer stability and long term storage.

Abstract: Methods using two to (n) purification columns to separate full length 5?-DMT-on oligonucleotides with size ranging from 40 to 180-mers from short length 5?-DMT-on oligonucleotides. Two of the said methods require using some columns sequentially with the collection and reprocessing of an intermediate fraction and are used for oligonucleotides with length ranging from 70 to 180-mers. A third method is carried out with columns stacked and used in series and is best used to purify oligonucleotides with length ranging from 40 to 80-mers. In the presence of a high ionic strength buffer, the short length DMT-on oligonucleotides bind to the top stacked columns while the less hydrophobic contaminant or DMT-off failures do not bind and/or are being washed off. In a stacked configuration, the full length DMT-on oligonucleotides are retained by the bottom column while in a 15 sequential configuration, full length DMT-on oligonucleotides are collected and reprocessed.

Abstract: Cylindrical devices (frits) are prepared by embedding aminoalkyl- or mercaptoalkyl-modified Controlled Pore Glass (CPG) in high-density polyethylene. Methods and devices pertaining to their use in the synthesis of nucleic acids are described. A reusable synthesis column or a reusable 96-chamber synthesis plate have been designed to hold one to 96 of the said frits that are inserted reproducibly into the synthesis chambers with a frit insertor. A short gas surpressure is required to drive entry of chemical reagents into the said frit. Reagents are retained into the frit until a second, longer surpressure is applied to drain the said reagents.

Abstract: Cylindrical devices (frits) are prepared by embedding aminoalkyl- or mercaptoalkyl-modified Controlled Pore Glass (CPG) in high-density polyethylene. Methods and devices pertaining to their use in the synthesis of nucleic acids are described. A reusable synthesis column or a reusable 96-chamber synthesis plate have been designed to hold one to 96 of the said frits that are inserted reproducibly into the synthesis chambers with a frit insertor. A short gas surpressure is required to drive entry of chemical reagents into the said frit. Reagents are retained into the frit until a second, longer surpressure is applied to drain the said reagents.

Abstract: The present invention relates to improved methods for the preparation of nucleic acids. More particularly, conventional solid supports used for nucleic acid synthesis are derivatized with activators having pKas within the 4 to 7 range. Preferentially, CPG-based solid supports are reacted with trialkoxysilanes containing an activator moiety such as pyridine. During each deblocking step of the nucleic acid synthesis cycle, bound pyridiniums are generated, yielding a weak acidic medium spreads throughout the solid support. The bound activators efficiently activate the phosphoramidite reagents towards coupling with 5?-hydroxynucleosides bound to the solid supports, thus eliminating or supplementing external deliveries of activator during the coupling steps.

Abstract: Cylindrical devices (frits) are prepared by embedding aminoalkyl- or mercaptoalkyl-modified Controlled Pore Glass (CPG) in high-density polyethylene. Methods and devices pertaining to their use in the synthesis of nucleic acids are described. A reusable synthesis column or a reusable 96-chamber synthesis plate have been designed to hold one to 96 of the said frits that are inserted reproducibly into the synthesis chambers with a frit insertor. A short gas surpressure is required to drive entry of chemical reagents into the said frit. Reagents are retained into the frit until a second, longer surpressure is applied to drain the said reagents.

Abstract: Herein, we introduce three multimodal SPE methods using two to (n) purification columns to separate full length 5?-DMT-on oligonucleotides with size ranging from 40 to 180-mers from short length 5?-DMT-on oligonucleotides. Two of the said methods require using some columns sequentially with the collection and reprocessing of an intermediate fraction and are used for oligonucleotides with length ranging from 70 to 180-mers. A third method is carried out with columns stacked and used in series and is best used to purify oligonucleotides with length ranging from 40 to 80-mers. Preferentially, a series of stacked columns contains from top to bottom hydrophobic porous sorbents with increasing pore sizes. Short length DMT-on oligonucleotides arise from depurination or branching during phosphoramidite based synthesis. Reversed phase partitioning and binding of short length DMT-on oligonucleotides take place simultaneously with the size exclusion of the full length DMT-on oligonucleotides.

Abstract: The present invention relates to improved methods for the preparation of nucleic acids. More particularly, conventional solid supports used for nucleic acid synthesis are derivatized with activators having pKas within the 4 to 7 range. Preferentially, CPG-based solid supports are reacted with trialkoxysilanes containing an activator moiety such as pyridine. During each deblocking step of the nucleic acid synthesis cycle, bound pyridiniums are generated, yielding a weak acidic medium spreads throughout the solid support. The bound activators efficiently activate the phosphoramidite reagents towards coupling with 5?-hydroxynucleosides bound to the solid supports, thus eliminating or supplementing external deliveries of activator during the coupling steps.

Abstract: The invention describes the synthesis of a panel of novel carbon-carbon linked carboranyl-containing 5,10,15,20-tetraphenylporphyrins bearing 25–44% boron by weight. In certain embodiments, a phenyl porphyrin compound has a carboranyl group attached to the phenyl group by a carbon-carbon linkage, wherein the phenyl group corresponds to the following formula where R7 through R11 are hydrogen, a carboranyl group, or are selected from the group consisting of hydroxyl, NMe3+, PMePh2+, PO(OH)2, SO3H, COOH, and NH2. In this embodiment, the carboranyl group is attached to the phenyl group by a carbon-carbon linkage, either one or two of R7 through R11 are other than hydrogen; and the phenyl porphyrin compound contains at least one phenyl group having at least one of said carboranyl groups.

Abstract: Polysubstituted catechol-based universal solid supports suitable for synthesizing oligonucleotides have been prepared. Following synthesis, cleavage of the oligonucleotide from the solid support and catechol-assisted elimination of the 3?-phosphate group is accomplished by treatment with standard basic media such as ammonium hydroxyde or1 methylamine.