Principle

Pre-filled single spin columns for dye-terminator removal

Unincorporated dye terminators will negatively affect analysis of sequencing results. Excess of dye terminators causes so-called "dye blobs" results in a partly unreadable sequence. NucleoSEQ® will remove unincorporated dye terminators. The subsequent analysis is of high quality with long reading length and minimized background. NucleoSEQ® columns are designed for fast, effective and cost efficient clean-up of sequencing reactions. The spin columns are prefilled with a dry size exclusion matrix which allows an efficient removal of dye terminators, for example BigDye™ terminators: the gel-filtration material consists of spheres with uniform pores and separates molecules according to molecular weight. After applying the seqencing reaction to the NucleoSEQ® column the small dye terminators and other impurities, for example salts, nucleotides, primers, traces of organic solvents are retained in the pores while labeled DNA fragments are excluded and recovered in the flow-through with high yield. In order to achieve long-time storage at room temperature NucleoSEQ® columns are prefilled with dry gel filtration resin. The matrix can easily be hydrated by adding water followed by an incubation period (> 30 min). Hydrated columns are ready-to-use and can be stored at 4°C for 14 days. A first short centrifugation step removes the remaining storage buffer. After loading the sample onto the column and a second centrifugation step, the DNA fragments of interest are recovered in the flow-through.

Features