FRET - Slide Preparation (Page 3)
At this time a second slide, also cleaned with a kimwipe, is placed on top of the taped slide. Scotch Tape is 62.5 nm thick and so the agarose forms a pad of similar thickness. The slide sandwich is then placed on a -20 C block that was just removed from a freezer and covered with a kimwipe. The agarose is allowed to harden on the block for between 50-60 seconds, depending on exactly how long the block is out of the freezer before one lays down the slide. The key is to harden the agarose, but not to let it freeze. If it freezes just throw the pad out, as freezing leads to high background in the images.
