High-throughput analysis of nucleic acid modification reactions using ion-pair reverse-phase high-performance liquid chromatography.
Ion-pair reverse-phase high-performance liquid chromatography is presented as a versatile platform for the rapid analysis of nucleic acid modification reactions in a high-throughput manner. This system allows both sensitive and nonradioactive assays to be developed for a variety of nucleic acid modification reactions. Examples presented here include assays for telomerase, uracil DNA glycosylase, polynucleotide kinase, T4 DNA ligase, C5-DNA methyltransferases, and the mismatch endonuclease CEL I. However, this approach is not confined to these reactions. Indeed the ability to perform a variety of nonradioactive assays with throughput times of 10 min per sample in conjunction with automated data analysis software represents a significant improvement in analytical and preparative nucleic acid enzymology.
Dickman
Mark J
MJ
Transgenomic Research Laboratory, Krebs Institute, University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, United Kingdom.
Matin
Maryam M
MM
Hornby
David P
DP
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D011088:DNA Ligases / Q000032:analysis* / Q000378:metabolism; D015254:DNA Modification Methylases / Q000032:analysis* / Q000378:metabolism; D004720:Endonucleases / Q000032:analysis* / Q000378:metabolism; D005453:Fluorescence; D009154:Mutation; D009696:Nucleic Acids / Q000378:metabolism; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000032:analysis* / Q000378:metabolism; D019098:Telomerase / Q000032:analysis* / Q000378:metabolism
D009696:Nucleic Acids; D015254:DNA Modification Methylases; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D019098:Telomerase; D004720:Endonucleases; C406922:CEL I nuclease; D011088:DNA Ligases