Fragmentation of RNA can be performed by enzymes, heat alone, or exposure to divalent cations at elevated temperature. This kit provides highly efficient and tunable RNA fragmentation by incubation with Magnesium ions at 94˚C.
The NEBNext® Magnesium RNA Fragmentation Module has been optimized to fragment RNA into small pieces by incubation with Magnesium ions at 94˚C. Fragment size is controlled by incubation time (Figure 1), and the reaction is ended by the addition of NEBNext Fragmentation Stop Solution.
Figure 1: Relative size distribution of eukaryotic mRNA fragments as seen using the Bioanalyzer 2100.
Poly(A)+ mRNA (40ng) purified from the Universal Human Reference Total RNA (Stratagene) was fragmented in 1 X NEB Next Magnesium RNA Fragmentation Buffer for 1–5 minutes at 94°C. Fragmentation Reaction was stopped in1X NEBNext Fragmentation Stop Solution. Samples were diluted 1:10 in Nuclease-Free Water and analyzed in the Bioanalyzer 2100.
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