From raw materials to validated system: The construction of a genomic library and microarray to interpret systemic perturbations in Northern bobwhite
The cDNA library was constructed from RNA pooled from Northern bobwhite brain, liver, testes, duodenum, colon and feather pulp tissues.  The SMART PCR cDNA Synthesis Kit (Clonetech Laboratories Inc. Mountain View, CA) was utilized to reverse-transcribe 0.5 µg of the poly(A) RNA into full length cDNAs.  The resulting cDNA library was examined using agarose-gel electrophoresis to examine the library for uniform abundance and distribution of cDNA size classes. The cDNA library was normalized prior to sequencing to capture both high and low abundance transcripts using the Trimmer cDNA Normalization Kit (Evrogen JSC, Moscow, Russia).