|
Maize Oligonucleotide Array Project |
|
Maize Oligonucleotide Array Protocols
Protocol Overview
The protocols listed below are those that are currently in use with the maize long-oligonucleotide arrays and have been optimized to be easy to use, produce optimal results, and be cost effective by using reduced amounts of reagent whenever possible.
We are currently recommending that Cy3/Cy5 labeled target be produced using RNA amplification as the protocol can accommodate small amounts of starting RNA (~1 ug total RNA) and produces enough target for multiple hybridizations should repeat hybridizations be necessary. An RNA isolation protocol that utilizes very small amounts of starting material is included that makes use of both Trizol and RNAeasy columns to produce ample amounts of very clean RNA that is ready for RNA amplification. The reduced amounts of reagents used in both of these protocols also means that they are cost effective.
Two hybridization protocols for cDNA and cRNA target hybridization are included. Both protocols give good results for cDNA and cRNA targets. The cDNA hybridization protocol is identical to the protocol that was on this site previously and is included for those array users who may have used it previously. We are currently recommending the cRNA hybridization protocol for cRNA targets as this protocol results in hybridizations with lower background and high signal to noise ratio. Please contact Dr. Roger Barthelson (rogerab@cals.arizona.edu) if you have any questions or comments regarding these protocols.
Last updated: April 1, 2008
RNA Isolation Using Trizol And Qiagen RNAeasy Columns (pdf, 29k)
Hybridization Protocol For cDNA Targets (pdf, 43k)
Hybridization Protocol For cRNA Targets (pdf, 38k)
cRNA Target Production Using RNA Amplification (pdf, 77k)
Last modified:April 1, 2008Thursday, 15-May-2008 09:10:31 MST