Fig. 1.
Wnt degenerate PCR. (A) Alignment of two highly conserved regions of Wnt proteins from Axolotl (aWnt), mouse (mWnt), human (hWnt), and Xenopus (xWnt). These regions are separated by 105 to 130 amino acids in various family members. Consensus sequence for the aligned protein regions is shown below with codon sequence. Oligonucleotides used for the degenerate PCR are shown in large arrows indicating orientation of the primers. (B) RT-PCR results of Wnt degenerate primers on RNA from FBMSC, FBM, 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 FBMSC, 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 reverse transcriptase was omitted from the reactions.