Wednesday April 16 notes
AMPure Bead Purification of yesterday’s double-digest samples.
- added 75 ul AMPure Beads to the 50 ul rxn. Incubated 5 min room temp
- 2 min on magnetic plate. removed solution
- added 200 ul 70% ethanol (fresh). removed. repeat
- off plate, added 40 ul H2O. 1 min
- on plate 2 min. transfered solution to new plate
Prep for ligation
- Remade P2 adapter as biotinylated DNA key for Streptavidin step
- 20 ul P2.1, 20 ul P2.2 (biotinylated), 10 ul 10x annealing buffer, 50 ul H2O
- anneal 97.5°C for 2.5, cool to 21°C by stepping down 3°C every minute
Meeting with SCH, NG, AN and MK to discuss thermal reactionome manuscript.