Nucleospin genomic dna from tissue kit

Nine Legionella strains (L. feeleii, L. anisa, L. bonzemanii, L. dumoffi, L. jordanis, L. longbeachae, L. micdadei, L. pneumophila segrogroup 1, L. pneumophila serogroup 2–14) were used in this study. All strains were cultured on Legionella selective medium GVPC at 37°C for 3–12 days. Other respiratory pathogens were cultured on their specific agars such as blood-agar media, MacConkey, cetrimide, chocolate and sabouraud agar. From Gram-negative bacteria, genomic DNA was extracted by heat shock (99°C for 20 min) and purified using the NucleoSpin^® Gel and PCR Clean-up extraction kit (Macherey-Nagel, Germany). Gram-positive bacteria and fungal pathogens genomic DNA were extracted using the NucleoSpin^® Genomic DNA from tissue kit (Macherey-Nagel, Germany), according to the manufacturer’s instructions. The genomic DNA was eluted with 100 μL of Milli-Q^® water and stored at −80°C until PCR assay.

We extracted gDNA from Apc^+/fl/Villin‐Cre^ERT2 and Apc^+/fl enteroids at D5 using the Nucleospin Genomic DNA from Tissue kit (Machery‐Nagel). The presence of the Apc‐mutated allele was detected by PCR using specific primers listed in Table S1.