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The 5'-end of the sagA/pel mRNA was determined by the extension of the 5'-end 32P-labeled oligonucleotides 5'-ACCTTATTTTAAAAATAAAGTTAA-3' following the method of Sambrook [ 45 ] . Oligonucleotides were labeled with [γ- 32P] ATP (10 mCi/mL in aqueous solution) (Amersham, Arlington Heights, IL) and T4 polynucleotide kinase (Gibco BRL Life Technologies, Rockville, MD).
The second chamber was purchased from GlycoTech (Product number 31-001, Rockville, MD).
Mouse nocturnin cDNA ( mNoc ) clones were obtained by screening a mouse BALB/c retinal cDNA library from ATCC (ATCC# 77448, Rockville, MD; reference [ 51]) using the standard protocol published in Sambrook, et al, 1989 [ 52] with probes from a human EST (T87026) purchased from Research Genetics, Incorporated (Huntsville, AL).
Rockville (MD): Department of Health and Human Services; 1995.
RPMI 1640 media, fetal bovine serum, and GlutaMAX™-I were obtained from Life Technologies (Rockville, MD).