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Researcher: I want to use an immobilized capture probe to isolate a specific DNA species. Which product is best suited for this purpose? |
TechTALKer: By covalently attaching the capture probe to a magnetic solid support, you obtain a stable, reusable system for isolating complementary sequences. The capture probe can be synthesized with a 5' amine-linker that provides a reactive site for covalent attachment to the solid support. MPG® Long Chain Alkylamine (LCA) is the particle of choice for covalently attaching primary amino-modified nucleic acids. The surface of the MPG® LCA magnetic porous glass particles are derivatized to provide reactive amine groups at the end of 15 Å spacer arms. These spacer arms enhance reactivity by minimizing steric hindrance. The amino groups, when reacted with the appropriate bifunctional reagent, can selectively bind to nucleic acids having a primary amino group. Amino-modified substrates used for coupling can be generated by either synthesizing oligonucleotides with an amino modifier phosphoramidite that incorporates an amino-linker at the 5' terminus, or incorporation of aminoallyl-dUTP or aminoallyl-UTP using DNA or RNA polymerase respectively. The oligo-coupled MPG® can be used for solid phase nucleic acid-based applications including solid phase RT-PCR; subtractive hybridization; purification of specific mRNA species; isolation of specific DNA sequences; hybridization-based assays; and purification of nucleic acid binding proteins. An easy to follow step-by-step procedure, Protocol 3.2 "Covalent Attachment of Oligonucleotides", is included with each purchase of MPG® LCA particles. This procedure provides instructions for covalent coupling of 5' amino-modified nucleic acids to MPG® LCA by reductive amination. We also offer this procedure in an optimized kit format. The Solid Phase Oligo COUPLE-IT™ Kit contains all the reagents and MPG® LCA solid support needed to carry out two efficient coupling reactions (10 mg MPG® LCA per reaction). The resulting product is an oligo-coupled magnetic solid support that can be stored for future use and, for many applications, is reusable.
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