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. 2005 Jan 13;33(1):e9. doi: 10.1093/nar/gni012

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© 2005, the authors Nucleic Acids Research, Vol. 33 No. 1 © Oxford University Press 2005; all rights reserved

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PCR amplification of bisulfite-treated human genomic DNA with primer pairs designed by the BiSearch software. The primers were used to amplify fragments of the human tyrosine hydroxylase gene. (A) Specific fragments of 195 and 262 bp were amplified at 56°C by primer pairs THbs113/THbas308 (lanes 1 and 2) and THbs565/THbas827 (lanes 3 and 4), respectively. The THbs565/THbas827 primer pair was also tested at 49°C annealing temperature. (B) Samples in lanes 1 and 2 are the same as in lanes 3 and 4 on panel A. The two other PCR products were obtained at the lower annealing temperature. The smears reveal an inefficient reaction. (C) The same gel after longer migration resolved the PCR products of lanes 3 and 4 into two distinct bands corresponding to the predicted lengths by BiSearch search.