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Table 1 Description of the release treatments methods applied to L. pneumophila strains and Acanthamoeba strains in a free-living state or in co-culture

From: Optimized methods for Legionella pneumophila release from its Acanthamoeba hosts

Release treatments methods Description Reference
Mechanical   
 21 gauge needle (21G) Passage three times through the needle  
 27 gauge needle (27G) Passage three times through the needle [22, 31]
 Centrifugation (Centrif.) Centrifugation at 10,000 × g for 10 min [21]
 Freeze in liquid nitrogen
 (N2)
Immerse liquid N2 for 2 min and thawed at 35 °C for 10 min [33]
 Freezing-thawing cycles
 (Freeze Thaw.)
3 cycles of freezing-thawing consisting of 15 min at −80 °C followed by 10 min at 35 °C [18]
Chemical   
 Triton™ X-100 0.02 % Triton™ X-100 for 20 min at RT [23]
 SDS 0.5 % SDS for 10 min at rt [24]
 Salt treatment (KCl) Centrifugation at 10,000 × g for 10 min, the supernatant was replaced with 1 mL of 0.038 M KCl, vortexed and incubated for 3 h at RT [19]