ARK-Genomics, Roslin Institute, Roslin, Midlothian, EH25 9PS, United Kingdom
Manufacture protocol
Oligos were resuspended in Corning Pronto Epoxide spotting solution to give a final concentration of 20uM. DNA was printed on Corning Epoxide slides using a Genomic Solutions MicroGrid II arrayer and the MicroSpot 10K pins, according to manufacturer's instructions. All printing was carried out in conditions of constant humidity (52% RH) and temperature (20C) in a room supplied with HEPA filtered air. Prior to printing the pins were cleaned by ultra-sonication in water and if necessary 50mM potassium hydroxide. A test print of all pins was carried out by printing Cy3 streptavidin conjugate dye in printing buffer onto epoxide slides and printing 100 features per pin. When all pins were printing consistent size spots for more than 80 features they were accepted as ready to print. The MicroGrid II was then programmed for the array print run, with a grid layout file prepared to allow duplicate printing atrandom within a patch. At the end of the print run the slides were removed following the program instructions in preparation for post processing. Slides dessicated overnight and then transferred to a humid chamber (45% humidity, 42C) for 8 hours. The slides were then washed in 0.2% SDS (room temperature) for 2 minutes with agitation before being rinsed in three water washes (30seconds, 30 seconds, 1 minute) with agitation. After these steps the slides were then incubated in water at 50C for 20 minutes before spin drying. The slides were then stored in a desiccator in the dark ready for use.
Less...
More...