Overcoming false negatives due to the genomic context in polymerase chain reaction amplification.
In some instances defined genomic regions are so poorly amplified that they seem to be unamplifiable. A protocol was developed which allows good PCR amplifications by the use of restriction digestion combined with the elution of a pool of restriction fragments of defined size range from agarose gel after electrophoresis. We describe the application of the method in the PCR amplification of a region of the white locus of Drosophila melanogaster that otherwise may be considered as a negative result.
Baldrich
E
E
Departament de Genètica i de Microbiologia, Universitat Autònoma de Barcelona, Spain.
Xamena
N
N
Cabré
O
O
D000818:Animals; D017931:DNA Primers; D004262:DNA Restriction Enzymes; D004331:Drosophila melanogaster / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D005188:False Negative Reactions; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004262:DNA Restriction Enzymes