Neurospora crassa Microarrays available from the FGSC

Nov 09, 2005

by UCB Neurospora Microarray Team

10,918 oligomers are on the chip.  10,526 oligos are based on MIPS or Broad Institute predicted ORFs.  8 oligos are Abmion ArrayControl Sense Oligo Spots (#1781).  The remaining 384 oligos (ChIP_Chip) were mainly designed in the intergenic regions.

40uM oligos in 3x SSC were printed onto Corning UltraGAPS slides.  Slides are not post-processed.  Rehydration is optional.  Oligos need to be cross-linked with 600mJ UV.

Slides should be stable for a year at room temperature in desiccator.

Slides may display unacceptable levels of green background and only way we know to remove is using the Pronto!™ Background Reduction Kit (Item #40029) from Corning.  It is not 100% effective so we recommend prescanning after prehybridization and identifying spots that are 3x standard deviation above background for possible elimination.

A standard protocol for reverse transcription and hybridization can be found here but we recommend to use Corning/Promega kit:

40076 Pronto Plus Indirect System

and RPN5661 Amersham Post-labeling Reactive Dye Pack.

which are expensive but always give brilliant results.

GAL files for spot gridding are batch dependant.  Please choose the appropriate one.

Slides                                                gal files

NC 6/2/05                              NC6_05.gal

NC_8/23/05 UCB                 NC8_05.gal

NC_10/26/05 CNR               NC9_05_CNR.gal

NC_10/04/05 UCB               NC10_05.gal

http://web.uconn.edu/townsend/Links/ffdatabase/downloads.html