Cetyltrimethylammonium bromide (CTAB) DNA extraction in plants is an effective and cost-efficient method, and therefore widely used across genetic studies. The versatility of CTAB has led to specialization and variation in DNA extraction methods; however, protocols often do not detail the modifications that are applied to optimize the extraction of DNA. To determine the effects of modifying extraction protocols on DNA output, experimental studies of each step of the extraction process are required. Our study tested alterations to the lysis step of CTAB extractions of silica-dried and herbarium specimens to systematically determine the effects of incubation temperature and duration across four representative angiosperm groups. Due to high levels of secondary metabolites in extractants of one species, we performed additional tests with chemical additives (sodium dodecyl sulfate [SDS], polyvinylpyrrolidone [PVP], and sorbitol) added during the lysis step. We evaluated the effects on extractant purity, DNA concentration, and fragment length, and found the changes in extractant purity were significant. Preservation method (silica or dried herbarium specimens) also significantly impacted DNA fragment length, purity, and yield, with silica-preserved tissue resulting in higher yields of unfragmented and clean DNA. Results from the additive experiment showed only PVP as significantly effective when measuring DNA yield compared to the other additives, and slightly more effective when compared to extractions using only CTAB. We recommend shorter and cooler lysis periods in CTAB plus PVP DNA extractions and using silica-preserved tissues due to the lower fragmentation, higher purity, and time effectiveness when compared to the hotter and longer lysis periods.