title: Analysis of mismatch repair in human nuclear extracts.
abstract: This unit describes an assay for completed mismatch correction along with a precise and sensitive assay for mismatch-repair excision intermediates and a simple and efficient procedure for construction of mismatch repair substrates. These substrates are derived by ligating synthetic oligomers into a gapped plasmid generated using the sequence-specific N.BstNBI nicking endonuclease, then the sequence-specific nicking endonuclease N.AlwI, to introduce single nicks for initiation of excision. These methods can be easily adapted for construction and analysis of DNA-lesion-containing substrates to study DNA-repair pathways other than mismatch repair.
authors.affiliation: Oregon State University, Corvallis, OR, USA.
authors.forename: Huixian
authors.initials: H
authors.lastname: Wang
authors.forename: John B
authors.initials: JB
authors.lastname: Hays
mesh_terms: D020137:Base Pair Mismatch; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism*; D017931:DNA Primers; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids
chemical_list: D017931:DNA Primers