Figure 1. Assessment of retinal crystallin gene copy number by quantitative RT-PCR

Four different mouse retinal samples were examined to determine the mean PCR cycle change from Hprt for each crystallin gene. Three pairs of retinas isolated from different mice (M1629, M2407, M2408) and a fourth sample consisting of 8 mouse retinas (WT-Pool) were analyzed. Total RNA was extracted from these samples and served as a template for reverse transcription. Primer pairs were designed to flank introns so PCR amplicons produced from a genomic DNA template could be detected. A cycle change of (+1) from Hprt represents a hypothetical two-fold increase in the mRNA abundance for that gene compared to Hprt mRNA expression levels. Every sequential increase of (+1) would represent a further two-fold increase in abundance of that specific transcript assuming 100% reaction efficiency in the RT-PCR. Data for α, β, γ, and other (μ, ζ, λ) crystallin gene members is are shown in panels A, B, C, and D, respectively.