Quantitative aspects of the mutant analysis by PCR and restriction enzyme cleavage (MAPREC).
Quantitation of virus revertants by PCR and restriction enzyme cleavage may give nonlinear results and, in some cases, produce artifacts caused by nucleotide misincorporation and heteroduplex formation, occurring during PCR. Modifications of the procedure allowed us to overcome these problems and develop a highly sensitive and reliable method of mutant quantitation. This procedure can be used to assess the quality of live vaccines and to study heterogeneity of viral and bacterial populations.
Lu
Z
Z
Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland 20892.
Douthitt
M P
MP
Taffs
R E
RE
Ran
Y
Y
Norwood
L P
LP
Chumakov
K M
KM
D001483:Base Sequence; D004252:DNA Mutational Analysis / Q000379:methods*; D017931:DNA Primers; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009154:Mutation; D009692:Nucleic Acid Heteroduplexes; D017955:Poliovirus / Q000235:genetics*; D011055:Poliovirus Vaccine, Oral / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D009692:Nucleic Acid Heteroduplexes; D011055:Poliovirus Vaccine, Oral; D004262:DNA Restriction Enzymes