Picture from Mars J et al. (2024) MicroPubl Biol "Low-cost, rapid and reliable DNA extraction from mammalian organoids using a ...."

Figure 1. Single Worm Lysis Buffer can be used to reliably extract DNA from mammalian organoids using a low-cost and rapid protocol: (A) Agarose gel electrophoresis of PCR products obtained using DNA extracted from two organoid lines using one of three indicated methods: Zymo Quick-DNA microprep kit, Biosearch Technologies' QuickExtract Solution or Single Worm Lysis Buffer. For mouse small intestinal organoids (M. musculus siOrgs), Eva1c PCR products are visible at the expected size of 1162 bp; for human fetal hepatocyte organoids (H. sapiens hepOrgs), E2F8 PCR products are visible at the expected size of 790 bp. For both organoid lines, either single organoids or 1 µl of organoid suspension was used and water controls were included for all experiments.(B) Agarose gel electrophoresis showing that 5-month storage at -20°C of a lysed M. musculus small intestinal organoid sample using Single Worm Lysis Buffer does not affect the stability of the extracted DNA. 1 µl of organoid suspension was used for DNA extraction; Eva1c PCR products are visible at the expected size of 1162 bp.(C) Agarose gel electrophoresis comparing different incubation times at 65°C and 95°C of the Single Worm Lysis Buffer protocol for M. musculus small intestinal organoid samples. 5 minutes at 65°C, followed by 2 minutes at 95°C is sufficient to prepare organoid lysate that will result in prominent, single bands after PCR. 1 µl of organoid suspension was used for DNA extraction; Eva1c PCR products are visible at the expected size of 1162 bp for all tested incubation times.All experiments were performed at least three times, the images show representative results.