Preparation of Adaptor Ligated DNA - NEBNext Fast DNA Fragmentation & Library Prep Set for Ion Torrent (E6285)
- Add the following to the microfuge tube:
For 10 ng For 100 ng – 1 μg Bst DNA Polymerase, Large Fragment 1 µl 1 µl Sterile H2O 9 µl 1 µl T4 DNA Ligase Buffer for Ion Torrent 4 µl 4 µl NEBNext DNALibrary Adaptors for Ion Torrent 2 µl 10 µl T4 DNA Ligase 4 µl 4 µl Total volume 20 µl 20 µl
- The total volume in the microfuge tube should be 40 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.
- Add 5 μl Stop Buffer, vortex and pulse-spin.