Pharmacia Biotech Inc.

Pharmacia P-l Biochemicals Inc.

North Carolina State University

James F Jolly

Suzanne B Treml

David W Walker

Rama P Ramanujam

Cristine J Dall

Robert A Luhm

Connie A Draveling

Robert M Kelly

Jack G Chirikjian

Phillip P Franciskovich

Brent A Burdick

Eve Y Ting

Chris R Lively

Olga B d'Hennezel

William A Riedl

Jean A Heaster

Susan G Frackman

Robert A DiFrancesco

Daniel D Zender

Caecilia J Huang

Guy D Duffaud

A reagent sphere is disclosed comprising at least one biological reagent and a glass-forming filler material in a concentration sufficient to facilitate formation of a glassy, porous composition, wherein the reagent semi-sphere is water soluble and has a T.sub.g sufficient for room temperature stability. At least two carbohydrates are combined to create the reagent sphere. The first carbohydrate is a synthetic high molecular weight polymer. The second carbohydrate is different than the first carbohydrate. A method is provided for making the reagent sphere.

Biological reagent spheres

An apparatus for dispensing reagent spheres from a vial includes a housing with an inlet through which the reagent spheres are received from the vial. Within the housing is a dispensing tube which forms a passageway for the reagent spheres and which has a first relatively large section in communication with the inlet and a funnel section in which the passageway reduces in cross section to a narrower linearizing section. The cross sectional area of the linearizing section is only slightly larger than one of the reagent spheres being dispensed. A metering shaft extends across and blocks the linearizing section of the passageway. The metering shaft has a depression for conveying one reagent sphere at a time between portions of linearizing section when the metering shaft is rotated 180 degrees. A gear on the metering shaft meshes with teeth on a plunger which is slidably mounted to the housing. A user pressing the plunger causes the metering shaft to rotate transferring a reagent sphere to a portion of the passageway connected to the outlet of the housing and thereby ejects that reagent sphere from the housing.

Dispenser for dried biological reagent spheres

A preparation of a purified DNA polymerase substantially identical to native DNA polymerase isolated from Thermococcus barossii is disclosed. In a preferable form of the present invention, the DNA polymerase is approximately 68 kDa in size, retains at least 90% activity after 8 hours of incubation at 95.degree. C. and retains between 60% and 80% proof-reading ability when catalyzing a polymerization reaction with a primer with a 3 base pair mismatch.

Purified Thermococcus barossii DNA polymerase

A reagent semi-sphere is disclosed comprising at least one biological reagent and a glass forming filler material in a concentration sufficient to facilitate formation of a glassy, porous composition, wherein the reagent semi-sphere is room temperature stable, water soluble, and has a T.sub.g above room temperature. A method is provided for making the reagent semi-sphere comprising providing an aqueous solution of a buffered biological reagent; mixing a glass forming filler material with the buffered reagent solution to form an emulsion wherein the concentration of the filler material is sufficient to facilitate formation of a glassy, porous composition having a predetermined semi-spherical shape; dispensing the emulsion in the form of substantially uniform droplets; collecting the droplets on an inert medium to form semi-spheres; and vacuum drying the droplets, under conditions suitable for maintaining the predetermined semi-spherical shape, to form the reagent semi-sphere; wherein the reagent semi-sphere is room temperature stable, water soluble, and has a T.sub.g above room temperature.

Room temperature stable reagent semi-spheres

A bacterial preparation capable of packaging phage .lambda. DNA is disclosed. This preparation is in lyophilized form and is stable at ambient temperature. In a preferred form, the preparation contains an over-expressed terminase protein, is prepared from the bacterial strain E. coli Cla [.lambda. c/857 Sam7 .DELTA.(cos-Nu1-A)::Kn.sup.r ]/.lambda. pTER and is capable of a packaging efficiency of at least 1.times.10.sup.8 pfu/.mu.g wild type .lambda. DNA. The present invention is also a method of creating a phage .lambda. DNA packaging extract comprising the steps of preparing a bacterial extract capable of packaging phage .lambda. and lyophilizing said extract.

In vitro phage lambda DNA packaging system

A room temperature stable restriction enzyme in a glassified carbohydrate stabilizer is disclosed. A restriction enzyme reaction buffer containing Mg.sup.+2 is dispersed in the stabilizer. One cleaves DNA merely by adding water and DNA to the composition. The composition can reduce glycerol-caused star activity by avoiding the need for glycerol. The composition also reduces star activity caused by high concentrations of enzymes. Another method is provided of comparing cleaved DNA samples in connection with forensic and paternity testing applications.

Glassified restriction enzymes

Growing community of inventors

Glendale, WI, United States of America

James F Jolly