Preparation of G-50 plates for cleaning up thermal cycling reactions:
Plates can be prepared well in advance of use, which is especially convenient since they take a minimum of 3 hours to prepare.
Using the black metal Millipore plate and plastic scraper made for this purpose add 45 μl of dry Sephadex G-50 to each well of a 96-well filter plate. [Please keep the black metal plate and scraper dry and do not allow the G-50 to get on the bench top. Return clean, unused G-50 to the stock bottle – it is expensive (about $500).]
To each well of the filter plate add 300 μl of distilled water (if adding water manually) or 290 μl of distilled water (if using the Hydra). [Do not use the Hydra unless you have received instructions and obtained permission.]
Place a clear plastic lid over the plate.
Allow the G-50 to hydrate for 3 hr at room temperature in a tightly sealed container containing a wet paper towel to keep a high moisture level, or preferentially overnight at 4°
C (in a tightly sealed container containing a wet paper towel to keep a high moisture level). Do not leave at room temperature longer than 3 hrs!!!!
If hydrated at room temperature, transfer to 4°
C until used.
Store hydrated plates at 4°
C inside a sealed container. Place a paper towel wetted with water inside the container to ensure that the G-50 remains fully hydrated.
Each time a plate is used, please write your initials and the date on the plate so that we can keep track of how often the plate has been used. Thanks