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Microarray Resource > Microarray Printing
Microarray PRINTING Arrays
are printed in a Class 100 cleanroom at 72oC, 55% RH, on to TeleChem Superclean (TeleChem
International Inc., Sunnyvale, CA) microscope slides coated "in house" with poly-L-lysine
according to Pat Brown protocol
in a Class 1000 cleanroom.
cDNA arrays are composed of PCR products generated from cDNA clones.
Plasmid DNA is generated using Qiagen TurboPreps on QiaRobot 9600
robots. PCR products are generated in 96-well plate format using
PE GeneAmp Thermocylers and are characterized on 1% agarose gels.
After assessing global plate DNA concentration, the products are
transferred to 384-well print plates using a Matrix Technologies Hydra with
Twister H. PCR products are
printed from 5 ml
sample volume at between 200 and 400 ng/ml in print
buffer (3XSSC, 0.005% sarcosyl) in Genetix X6004 plates. Duplicate adjacent spots
are printed when possible using TeleChem Stealth SMP3 split pins on a
GeneMachines Omnigrid-100 arrayer. Features are printed at 175
mm center-to-center
distance using 16, 32 or 48 pins in a 4 x 4, 4 x 8 or 4 x 12 subarray pattern with
up to 25 rows and 25 columns
per subarray. Oligonucleotide arrays are composed of Operon or Compugen oligos
- printed, single spot, at 40 mM in print buffer (3XSSC, 0.01% sarcosyl )
as above. All arrays for standard fluorescence are outlined at
the corners by
scoring with a diamond pencil on the reverse side of the slide. Slides printed
for Genicon Resonance Light Scattering applications are not scored. All slides are labeled with slide
type and a unique number on a Brothers white tape and are
post-processed according to
Pat Brown protocol
before storing at room temperature in plastic slide boxes (no cork or
paper inserts) inside plexiglass cabinets with desiccant until ready for use. |
