Stratagene

About: Stratagene is a based out in . It is known for research contribution in the topics: Nucleic acid & Gene. The organization has 161 authors who have published 232 publications receiving 17223 citations.

Small-sample total RNA purification: laser capture microdissection and cultured cell applications

Abstract: Gene expression studies require analysis of RNA, but isolation of total RNA from very small samples by traditional methods can be difficult and inefficient. The Absolutely RNA microprep kit provides a convenient method for isolating total RNA from small numbers of cells such as those harvested by laser capture microdissection (LCM). The protocol includes binding of RNA to a solid support, thus eliminating the need for organic extraction and alcohol precipitation. DNase digestion on the solid support reduces or eliminates DNA contamination and minimizes RNA handling. Efficient washing removes contaminants, and elution in a small volume of buffer results in high-purity RNA at a concentration appropriate for demanding applications such as RT-PCR. RNA isolated from as few as 200 laser capture microdissected brain tumor cells resulted in detection of low, medium, and highly expressed genes by conventional and real-time RT-PCR.

Affinity purification of recombinant proteins fused to calmodulin or to calmodulin-binding peptides.

Abstract: Publisher Summary The interaction between calmodulin (CaM) and its various ligands has been exploited for the affinity purification of recombinant proteins from Escherichia coli extracts in a number of systems. CaM is a calcium-binding protein that plays a central role in regulating a wide range of calcium dependent intracellular processes. This chapter focuses on the purification of fusion proteins containing the 26 amino acid calmodulin-binding peptide (CBP) derived from the C terminus of skeletal muscle myosin light-chain kinase using a solid support resin containing covalently bound calmodulin (CaM). Several alternative systems in which CaM is employed as a fusion partner are also described in the chapter. The high degree of specificity of CBP for CaM together with the stringent wash conditions, allowed because of the strength of the CBP-CaM interaction, results in the consistent recovery of highly purified CBP fusions. The small size of the peptide ( E. coli , and thus a wide range of proteins are consistently expressed to high levels when the CBP is positioned at the N terminus of the fusion.

Methods of enriching for and identifying polymorphisms

Abstract: The invention encompasses methods for enriching for and identifying a polymorphism within a nucleic acid sample either by separating a subset of a nucleic acid sample or by selectively replicating a subset of a nucleic acid sample such that the polymorphism is contained within a nucleic acid population with reduced complexity, and then identifying the polymorphism within the enriched nucleic acid sample. Methods also are disclosed for enriching for and identifying a polymorphism by contacting a nucleic acid sample that includes a subset of nucleic acid molecules having a sequence that binds to a sequence-specific binding activity with a molecule having a sequence-specific binding activity under conditions which permit specific binding, such that the subset of nucleic acid molecules bound to the activity is enriched for nucleic acid molecules having the sequence recognized by the sequence-specific binding activity, and detecting a polymorphism with respect to a reference sequence in the subset of nucleic acid molecules.

Method and apparatus for cover assembly for thermal cycling of samples

Abstract: A flexible heating cover assembly for an apparatus for heating samples of biological material with substantial temperature uniformity includes a housing having a plurality of engageable enclosure components; a resistive heater having a plurality of heater element areas; a heater backing plate providing stability to the resistive heater; a force distribution system that distributes a force over the heater backing plate; and a support plate providing stiffness for the force distribution system, wherein the arrangement of the resistive heater, the heater backing plate, the force distribution system and the support plate provide substantial temperature uniformity among a plurality of sample tubes for receiving samples of biological material. The flexible heating cover assembly improves the uniformity, efficiency, quality, reliability and controllability of the thermal response during thermal cycling of the biological material.