Abstract
Objective and Method – 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 •–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. Conclusion – 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.
Publication Date
1-31-2000
First Page
11
Last Page
14
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Recommended Citation
M, Simsek and H, Adnan
(2000)
"Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction,"
Sultan Qaboos University Medical Journal: Vol. 2: 11-14.
DOI: https://doi.org/10.18295/2075-0528.2141
