An efficient laboratory protocol for the sequencing of large numbers of lacI mutants recovered from Big Blue transgenic animals.
Mutational spectra provide a powerful approach to investigate both the mutagenic potential and the mechanism of action of suspected mutagens and carcinogens. Recently, transgenic techniques have made it possible to generate mutational spectra in animals. Such a spectrum may consist of 50 to 200 mutants depending on the nature of the mutations, and many spectra can be generated depending on the design of the experiment. This report describes a practical approach for the processing and sequencing of large numbers of lacI mutants recovered from Big Blue animals.
Erfle
H L
HL
Biology Department, University of Victoria, British Columbia, Canada.
Walsh
D F
DF
Holcroft
J
J
Hague
N
N
de Boer
J G
JG
Glickman
B W
BW
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics*; D003205:Computing Methodologies; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D051379:Mice; D008822:Mice, Transgenic; D009154:Mutation; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D012097:Repressor Proteins