title: Effect of single mismatches at 3'-end of primers on polymerase chain reaction.
abstract: OBJECTIVE
  To investigate the effect of three different mismatches (G/T, G/A or G/G) at the 3'-end of a primer to amplify a 268 bp (base pair) region of the human β-globin gene using different annealing temperatures (45 to 65°C).
  
  
  RESULTS
  The primer with the G/T mismatch was as efficient as the normal primer (G/C match) in the amplification of a 268 bp product at all temperatures tested. However, the primers having G/A or G/G mismatches at the 3'-end did not produce any specific polymerase chain reaction (PCR) fragment at all the annealing temperatures used, except a barely detectable 268 bp product for the G/G mismatch at 45 and 50°C.
  
  
  CONCLUSIONS
  We conclude that our PCR system was refractory to amplification when one of the primers contained a G/A or G/G mismatch at the 3'-end with template DNA.
authors.affiliation: Department of Biochemistry, Sultan Qaboos University, P.O.Box: 35, Postal Code: 123, Muscat, Sultanate of Oman.
authors.forename: M
authors.initials: M
authors.lastname: Simsek
authors.forename: H
authors.initials: H
authors.lastname: Adnan
keywords: PCR; mismatched primers; β–globin gene
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