- Gel Purification: CMV/V0120, MV9
Using Prior Miniprep Digests
- CMV culture 1 - check with E/P
- MV9 culture 2 - check with P/X
- Used previously digested CMV and MV9.
- Placed 15μL 1kB Ladder in well one and 30μL of both CMV and MV9 digest in well two and three respectively.
- Ran gel at 110V for 30 minutes.
- Placed gel on UV transilluminator and cut out desired bands.
15 μL/lane; 1% agarose; Ladder
- Used Zymo gel purification kit.
- Assumed 200mg gel, added 600μL ADB to each 1.5mL tube.
- Put on heat block at 55°C for 5 minutes, vortexed and then placed back on the heat block for an additional 5 minutes, vortexed again.
- Put ~750μL gel/ADB into spin column, centrifuged at max rpm for 30 seconds.
- Pipetted 200μL wash buffer to each tube, centrifuged at max rpm for 30 seconds. Repeated once.
- Transferred columns to new labeled 1.5mL tubes.
- Pipetted 15μL elution buffer to each column.
- Spun at max rpm for 30 seconds.
- Put marked CMV and MV9 in -20°C fridge in box labeled Cameron.