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Microarray Resource
> Labeling and Hybridization
LABELING AND HYBRIDIZATION
The Resource provides labeling and
hybridization services for microarray projects ranging from pilot
studies to full blown projects with multiple parameters and multiple
replicates. Please contact
Irina Tikhonova to discuss your individual requirements.
LABELING STRATEGIES
We routinely use and
recommend the following labeling approaches that
do not involve nucleic acid amplification steps. Please follow
the links for detailed protocols optimized for slides fabricated at the
Keck Resource.
Amino-allyl
labeling
and monofunctional Cy-dye conjugation.
20-100
mg
total RNA; 5-10 mg
mRNA.
Genisphere
Dendrimer
labeling - Cy-dye.
2 mg
total RNA; 250 ng mRNA.
HYBRIDIZATION STRATEGIES
Two main approaches are
used in the Resource.
GeneMachines Hybridization Chambers -
used for manual hybs of arrays
with small dimensions where an 18 x 20 mm coverslip is sufficient to
cover the array.
Advalytix SlideBooster Autohyb Instrument
-
used for all 4 x 12 subarray
slides where a 60 x 22 mm coverslip would be needed for manual hyb. The
ArrayBooster works on the principle of Surface Accoustic Wave
technology to actively mix the liquid evenly over the array.
PROTOCOLS |
