This mix ensures that the DNA is firmly attached to the substrate.
Before you flush the mixes through a channel, calculate which dilution you are going to make (fill in the ? in the table below). Dilute the surface treatment mix components (BSA, neutravidin, Holliday Junction) in T50.
Insert the mixes in the right order, let them settle and remove the unbound part with flushing with T50 (as described).
* Please wrap the HJ in aluminum foil, to avoid photo-bleaching. If possible store it in the fridge.
Even if it dries out, you should be able to use several days later with new imaging buffer. The fluorescent intensity will have dropped.
Imaging buffer is required to allow your sample to be imaged for longer time.
Before making the imaging buffer, calculate which volume you are going to use (fill in the ? in the table below) in order to get to the right final concentration.
Gloxy should be added as last, just before the measurement starts!!
You can make the imaging buffer without gloxy in advance, and just before flushing the imaging buffer through the channel add the gloxy.
Gloxy will quickly drop the pH of your buffer, you can use buffer with gloxy for maybe an hour. After that, trash the remaining buffer with gloxy.
Use 100 µl of imaging buffer per channel.