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Our work highlights the importance of an iterative approach to mutagenesis, proving that large rate enhancements are achieved when mutations are made in already active mutants.
Chromosomal gene replacements can be constructed using double-stranded DNA (dsDNA) substrates with >500 bp of homology[27], and point mutations are made using single-stranded DNA (ssDNA) substrates with >45 bp homology[28].
The DNA damage checkpoint specifically delays entry into mitosis in the presence of damaged DNA to provide cells enough time to repair damaged DNA before permanent mutations are made [1].
2) Claims about effects of mutations are made without appropriate statistical analysis and rationale.
To elucidate the detailed catalytic mechanism of AHL lactonase, mutations are made on residues that presumably contribute to substrate binding and catalysis.
In the first place, when large-scale mutations are made less frequent relative to point mutations, the genome expansion is less pronounced and the success rate is lower.
Similar(51)
Mutations were made using PCR, and the mutated fragment was subcloned back into pCS2-Nrp1 using endogenous restriction sites.
The mutated 3'-UTR of CCND2 was synthesized after point mutations were made in several bases within the binding sites of these PCR products.
Mutations were made for each enzyme in the vicinity of the active site and we examined these variants for glycosylase and lyase activity.
To study the role of the potential hydrogen bonds that connect the two domains, a series of mutations were made to the hydrogen bond donors and acceptors along the domain interface.
Mutations were made and verified with sequencing.
More suggestions(15)
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