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-No Derivative Works 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
