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All subsequent steps followed the polyclonal and MAb protocols supplied with the Dako Envision+ HRP system.
This methodology was followed to simulate the results users would achieve by following current protocols supplied with each microarray system.
The Seleno-Met site was determined and refined using the protocols supplied with the program CNS using a dataset measured at three wavelengths (inflection, peak and remote).
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Reverse transcription reactions were carried out with 5 μg of total RNA following the standard protocol supplied with the reverse transcriptase.
Total GPx activity in the homogenate was measured according to the protocol supplied with the kit (Cayman Chemical, Ann Arbor, MI, USA).
For the Genome Analyzer, library constructions followed the protocol supplied with the Genomic DNA Sample Prep Kit (Illumina, "Preparing Samples for Sequencing Genomic DNA", version 1003806_Rev. B, March 2008) with minor modifications.
DNase treatment was performed to remove residual DNA from the RNA samples by use of the recombinant DNase I as specified in the protocol supplied with the Ambion DNA-free Kit (Applied Biosystems, Foster City, CA).
Total RNA was extracted using the protocol supplied with the Trizol reagent.
The resulting double stranded cDNA was purified by using column-based reagents, according to the protocol supplied with the kit.
The samples were extracted following the exact instructions in the protocol supplied with the kit (step 9 to 17).
This draft protocol, supplied with the reasoning as mentioned above, was sent to all participants of the Delphi-procedure.
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