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).
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.
Once you have imaging buffer, you need to add the Cy3 oligos, in order to get a fluorescent signal from the ends of the DNA