In vitro mutagenesis {site-directed mutagenesis} can study binding sites and functional regions. Site-directed mutagenesis hybridizes synthetic 10-base to 15-base oligonucleotides to DNA sites. Oligonucleotides differ from original sequences by one nucleotide at end. Oligonucleotides hybridize well to original sequences, because they differ by only one nucleotide. Hybridized sequences replicate to make mutated genes.
enzymes
DNA ligase connects perfectly aligned DNA strands. Mutated ends do not ligate {ligase-mediated}, showing mutation locations. RNase A cuts DNA-RNA complexes where sequences mismatch and can detect mutation locations. Osmium tetroxide and hydroxlamine cut at unmatched C or T bases. Restriction enzymes fragment single-strand DNA. Different DNA fragments have different conformations and so different mobilities {single-stranded conformation polymorphism} (SSCP).
Biological Sciences>Genetics>Mutation>Experiments
4-Genetics-Mutation-Experiments
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Date Modified: 2022.0224