Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction
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 efficie...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Sultan Qaboos University
2000-01-01
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| Series: | Sultan Qaboos University Medical Journal |
| Subjects: | |
| Online Access: | https://journals.squ.edu.om/index.php/squmj/article/view/1188 |
| _version_ | 1818001523635912704 |
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| author | Simsek M Adnan H |
| author_facet | Simsek M Adnan H |
| author_sort | Simsek M |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-14T03:35:24Z |
| format | Article |
| id | doaj.art-1cf0e0f9273c4739bd6e106e97d8cc1f |
| institution | Directory Open Access Journal |
| issn | 2075-051X 2075-0528 |
| language | English |
| last_indexed | 2024-04-14T03:35:24Z |
| publishDate | 2000-01-01 |
| publisher | Sultan Qaboos University |
| record_format | Article |
| series | Sultan Qaboos University Medical Journal |
| spelling | doaj.art-1cf0e0f9273c4739bd6e106e97d8cc1f2022-12-22T02:14:47ZengSultan Qaboos UniversitySultan Qaboos University Medical Journal2075-051X2075-05282000-01-012111141117Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain ReactionSimsek M0Adnan H1Department of Biochemistry, Sultan Qaboos University, P.O.Box: 35, Postal Code: 123, Muscat, Sultanate of OmanDepartment of Biochemistry, Sultan Qaboos University, P.O.Box: 35, Postal Code: 123, Muscat, Sultanate of OmanObjective 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.https://journals.squ.edu.om/index.php/squmj/article/view/1188pcr, mismatched primers, β−globin gene |
| spellingShingle | Simsek M Adnan H Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction Sultan Qaboos University Medical Journal pcr, mismatched primers, β−globin gene |
| title | Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction |
| title_full | Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction |
| title_fullStr | Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction |
| title_full_unstemmed | Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction |
| title_short | Effect of Single Mismatches at 3′–end of Primers on Polymerase Chain Reaction |
| title_sort | effect of single mismatches at 3 end of primers on polymerase chain reaction |
| topic | pcr, mismatched primers, β−globin gene |
| url | https://journals.squ.edu.om/index.php/squmj/article/view/1188 |
| work_keys_str_mv | AT simsekm effectofsinglemismatchesat3endofprimersonpolymerasechainreaction AT adnanh effectofsinglemismatchesat3endofprimersonpolymerasechainreaction |