Frizzled degenerate PCR. (A) Alignment of two highly conserved regions of fz proteins from Drosophila(Dfz), human (Hfz), mouse (Mfz), and rat (Rfz). These regions are separated by 60 to 90 amino acids in various family members. Consensus sequence for the aligned protein regions is shown below with corresponding codon sequence. Oligonucleotides used for the degenerate PCR are shown in large arrows indicating orientation of the primers. (B) RT-PCR results of fz degenerate primers on RNA from CD34⁺ FBM, CD34⁺ ABM, and a no-template control (−). Reverse transcription was performed with (+RT) or without (−RT) RT in the cDNA synthesis step. Equivalent amounts of RNA were used from each tissue for reverse transcription. One quarter of the cDNA was used for PCR amplification with the degenerate primers. (C) RT-PCR results using primers to GAPD. One quarter of the cDNA synthesized from FBM and ABM was PCR amplified using the GAPD primers. No product was observed in (B) and (C) for the no-template control or when RT was omitted from the reactions.