Abstract: The invention provides methods for improving the physical and chemical stabilities of therapeutic proteins in solutions by reducing or eliminating protein degradation, aggregation and precipitation using small molecule stabilizing agents such as proline, arginine and/or compounds capable of forming a Schiff bond with amino groups of the protein. The invention further provides liquid formulations of a monomeric insulin stabilized by one of more stabilizing agents such as proline, arginine and/or acetone. Also provided are methods of making and using the stabilized monomeric insulin formulations.

Abstract: A method for removing extraneous nucleotides in a cloned coding sequence using a type IIs endonuclease, the method comprising introducing a linker that comprises at least one recognition site for a Type IIs restriction endonuclease to the ORF, cloning the ORF into a suitable vector, and removing the extraneous nucleotides from the vector with a RE IIs digestion. Also provided are a vector, a kit and oligonucloetides suitable for the invention.

Abstract: Recombinant DNA vectors and methods for cloning and expressing nucleic acid molecules by using a combination of site-specific recombination and end-to-end joining or linking of nucleic acid molecules, such as endonuclease restriction digestion and ligation. The DNAs, vectors and methods can be used for inserting, exchanging, transferring a variety of DNA segment(s) both in vitro and in vivo. Also disclosed are linker molecules and methods using these linkers the can be used for cloning a gene of interest into an expression vector in one-step. The linker sequences comprise adapter sequences for cloning purposes, as well as eukaryotic and prokaryotic ribosome binding sites for increase translation efficiency.

Abstract: The sensitivity and specificity of polynucleotide synthesis is increased by protecting the 3′-end of an oligonucleotide used as a primer in the synthesis of the polynucleotide. Protection of the 3′-end of an oligonucleotide prevents non-specific chain elongation. Removal of blocking group an elevated temperature, using a thermostable enzyme, permits template-specific polynucleotide synthesis. The present invention also provides oligonucleotides with a 3′ end protected by a blocking group and a thermostable enzyme capable of removing the blocking group at an elevated temperature. The compositions and methods of the invention are very useful in a variety of techniques for DNA/RNA amplification and analysis, including medical genetics research and diagnosis, pathogen detection, forensic, and animal and plant genetics applications, among others.

Abstract: The sensitivity and specificity of polynucleotide synthesis is increased by protecting the 3′-end of an oligonucleotide used as a primer in the synthesis of the polynucleotide. Protection of the 3′-end of an oligonucleotide prevents non-specific chain elongation. Removal of blocking group an elevated temperature, using a thermostable enzyme, permits template-specific polynucleotide synthesis. The present invention also provides oligonucleotides with a 3′ end protected by a blocking group and a thermostable enzyme capable of removing the blocking group at an elevated temperature. The compositions and methods of the invention are very useful in a variety of techniques for DNA/RNA amplification and analysis, including medical genetics research and diagnosis, pathogen detection, forensic, and animal and plant genetics applications, among others.