Novagen, Inc.

Pel-freez Rabbit Meat, Inc.

Emd Biosciences, Inc.

Robert C Mierendorf

Robert E Novy

Thomas R Van Oosbree

Gary L Milburn

Mark McCormick

David O'Reilly

Richard Garber

Barbara B Morris

Kristin M Kolb

Yuping Ambuel

Beth Hammer

There is now a market for kits containing the necessary components for performing protein synthesis reactions using template DNA of the experimenter. One category of existing system for performing protein synthesis reactions is based on an S-30 extract from the bacteria. It is disclosed here that a simple fractionation process can dramatically improve the performance of an S-30 prokaryotic protein synthesis reaction mixture. In one embodiment, the fractionation is a simple freezing and thawing of an S-30 extract combined with a supplemental mix, followed by centrifugation. The resulting fractionated S-30 reaction mixture yields more full-length target protein and less non-full length or non-target protein than possible using prior art S-30 systems.

E. coli extract for protein synthesis

An antibody raised against an epitope of a peptide having an amino acid sequence selected from a group consisting of Leu Asp Gly Tyr Pro Ser Asp Tyr Asp Pro Phe Val Glu Asn Ala Gln Ile Tyr (SEQ ID NO:1) and Leu Asp Gly Tyr Pro Ser Asp Tyr Asp Pro Phe Glu Glu Asn Ala Gln Ile Tyr (SEQ ID NO:2) is advantageously used in an indirect immunoassay to detect the presence of active Human Herpesvirus-6 in a patient sample.

Antibody to carboxy-terminus of human herpesvirus 6 immediate early

A method for performing coupled in vitro transcription and translation reactions is disclosed. In the transcription reaction, the quantity of mRNA produced is limited to a level of less than about 2.5 micrograms in a 10 microliter volume prior to the translation elements being added. Limiting the level of mRNA produced prevents saturation of the translational processes and thus aids in the efficiency and fidelity of the translation process.

Method for in vitro protein synthesis

A method for performing coupled in vitro transcription and translation reactions is disclosed In the transcription reaction, the quantity of mRNA produced is limited to a level of less than about 2.5 micrograms in a 10 microliter volume prior to the translation elements being added. Limiting the level of mRNA produced prevents saturation of the translational processes and thus aids in the efficiency and fidelity of the translation process.

A vector for the direct cloning of the products of PCR protocol incorporates single nucleotide overhangs at one or both ends of a linearized DNA segment. The single nucleotide overhangs are uracil or inosine residues, as desired, to facilitate cloning of the desired PCR products.

Direct cloning of DNA fragments

An antibody raised against an epitope of a peptide having an amino acid sequence selected from a group consisting of LDGYPSDYDPFVENAQIY (SEQ ID NO:1) and LDGYPSDYDPFEENAQIY (SEQ ID NO:2) is advantageously used in an indirect immunoassay to detect the presence of active Human Herpesvirus-6 in a patient sample.

Human herpesvirus 6 detection assay

A method for forming directionally clonable randomly primed cDNA molecules includes the step of priming first strand synthesis using a set of first strand cDNA primers having the sequence 5'-XXNNNNNN-3' where XX is a constant dinucleotide pair across the set and NNNNNN is a random hexanucleotide, the set including primers representing all random hexanucleotides.

Method and kit for making CDNA library

A method for determining the nucleic acid sequence that encodes ligand binding domain on any protein encoded by a known gene includes the steps of non-specifically cleaving the coding region of the known gene, size-selecting the cleavage products, cloning the size-selected gene products into an expression vector active in a heterologous host, using a novel cloning strategy expressing the peptides produced by the clones and screening for clones that produce peptides that interact with an antibody or ligand of interest.

Growing community of inventors

Madison, WI, United States of America

Robert C Mierendorf