Preparation of Adaptor Ligated DNA - NEBNext Fast DNA Library Prep Set for Ion Torrent (E6270)
- Add the following to the microfuge tube:
For 10 ng For 100 ng – 1 µg Sterile H2O 18 µl 3 µl T4 DNA Ligase Buffer for Ion Torrent 10 µl 10 µl Bst 2.0 WarmStart DNA Polymerase 1 µl 1 µl NEBNext DNA Library Adaptors for Ion Torrent 5 µl 20 µl T4 DNA Ligase 6 µl 6 µl Total volume 40 µl 40 µl
- The total volume in the microfuge tube should be 100 µl. Mix the contents by pipetting up and down several times.
- Incubate in a thermal cycler for 15 minutes at 25°C, followed by 5 minutes at 65°C, hold at 4°C.
If performing size selection with beads, proceed directly to size selection using AMPure XP Beads. If using E-gel or agarose gel for size selection, proceed to Cleanup of Adaptor Ligated DNA before proceeding to gel-based size selection.