Evaluation and Modification of Plasmid DNA Purification Kits

摘要

Rapid polymerase chain reaction (PCR) amplification of DNA in small volumes requires more stringent controls of DNA purity. Although DNA extracted using commercial kits meets purity requirements, they are time consuming and inefficient Kit protocols were modified to not only remove PCR inhibitors, but also to increase efficiency. Using modified commercial plasmid purification kits, we were able to amplify plasmid DNA extracted from field samples containing humic acid, a well known PCR inhibitor. The DNA extracted was amplified using the LightCycler, a hot air thermal cycler that uses cycle times of <1 min/cycle and provides real time detection of target material using dsDNA dye intercalation followed by melting curve analysis.