Figure 5.

Analysis of the tissue distribution of V(D)J RAG expression. Total RNA prepared from different tissues of a 4-week-old chicken was subjected to DNase treatment before synthesizing cDNA by using an oligo(dT)₍₁₅₎ primer and avian myeloblastosis virus reverse transcriptase. PCR products obtained with RAG-1 and RAG-2 specific primers (Table 1) were separated in 1% agarose gels, transferred onto nitrocellulose membranes, and hybridized with ³²P-labeled internal oligonucleotides. Genomic DNA was used as a positive PCR control. Membranes being examined for control actin products were exposed to x-ray films for 12 hr, whereas a 24-hr exposure time was used for the evaluation of RAG-1 and RAG-2 expression.