MicroSpin G-50 columns are designed for the rapid purification of DNA in desalting, buffer exchange, removal of dye terminators from cycle sequencing reactions and removal of labelled nucleotides from DNA labelling reactions.
For rapid purification of labeled DNA (DNA probes of > 20 bases) from unincorporated labeled nucleotides in a volume of 25 to 50 μL using spin-column chromatography.
Columns are prepacked with Sephadex G-50 DNA Grade pre-equilibrated in TE buffer with 0.05% Kathon CG/ICP Biocide.
Product recovery will be at least 10% lower than that from ProbeQuant G-50 Micro Columns.
Can also be used for buffer exchange/desalting of enzymatic reactions (10 to 100 μl) following heat denaturation and phenol extraction.
MicroSpin G-50 Columns, rapid purification of DNA requiring clean-up
Good product yield and purity is obtained with sample volumes from 12-50 μL. They are suitable for any DNA greater than 20 bases in length and will not remove or denature enzymes.
The resin used in these columns, Sephadex G-50 DNA Grade, can be used for a wide range of applications, including desalting DNA, buffer exchange, and removal of unincorporated nucleotides from end-labelled oligonucleotides.
Sephadex G-50 is one of five different G-types ranging from G-10 for small molecules to G-75 for larger molecules. Sephadex G-50 is a well-established size exclusion resin for desalting and buffer exchange of biomolecules > 30 000 molecular weight, and with a spin protocol can be used for DNA and oligo purification of molecules greater than 20 bases in length.
Alternatively, Sephadex G-25 or Sephadex G-100 can be used. Sephadex G-25 has an exclusion limit of approximately Mr 5000, and with a spin protocol this means it can be used for any DNA greater than 10 bases in length.
Sephadex G-100 DNA Grade has an exclusion limit of 25bp ds DNA.
All these Sephadex types are therefore suitable for the purification of oligonucleotides or small DNA fragments following synthesis or a labeling reaction.
