Bacterial isolation, genome amplification and DNA extraction

• Isolate ant host tissue or collect fecal droplets
• Suspend isolated material in 1000 µL cold SPG Buffer (218 mM sucrose, 3.8 mM KH₂PO₄, 7.2 mM K₂HPO₄, 4.9 mM l-glutamate, pH 7.2) or homogenize it (in cold SPG buffer) using an ice cold Wheaton glass homogenizer
• Centrifuge twice at 4 ˚C for 15 min at 3,200 g
• Pass the supernatant through a 5 µm (Acrodisc) syringe filter first, then a 2.7 µm (Whatman) syringe filter, and finally through a 1.3 µm (Acrodisc) syringe filter.
• Centrifuge  at 4 ˚C for 20 min at 18,000 g
• Resuspend the pellet (bacterial cells) in 5 µl SPG buffer
• Use 1 µl  for Multiple Displacement Amplification (MDA) to obtain whole genomic DNA using the Qiagen REPLI-g Midi Kit following the manufacturer’s instructions

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