Abstract
We present efficient and reproducible parallel strategies for preparing large quantities of pure heteroduplex plasmids containing defined mismatches. The strategies described involve the use of synthetic oligonucleotides, the commercially available pGEM-T plasmid, and nicking enzymes to prepare prerequisite ssDNA. Alternatively, bacterial packaging cell lines containing an engineered phagemid construct to produce ssDNA without the need of a helper phage were utilized, hence providing added flexibility and choice. These integrated approaches help to construct different mismatch substrates of choice in large quantities, thus enhancing the usability of mismatch repair assays and extending their range and accessibility to wider research groups.
Item Type: | Journal article |
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Faculties: | Geosciences > Department of Earth and Environmental Sciences |
Subjects: | 500 Science > 550 Earth sciences and geology |
ISSN: | 0003-2697 |
Language: | English |
Item ID: | 67905 |
Date Deposited: | 19. Jul 2019, 12:23 |
Last Modified: | 04. Nov 2020, 13:50 |