Isolating thermophilic bacteria - (Apr/26/2011 )
I'm wondering if any of you have experience isolating and identifying thermophilic bacteria. I'm trying to isolate different DNA ligases and am too cheap to pay ATCC
You might try pcr using a pcr enzyme commercially available as a template, if you know the sequence. There is contaminating genomic DNA in most enzyme preps. This works for restriction enzymes and methylases, as well.
I would plate a sample where thermophilic bacteria are expected to occur on a petri dish with standard media for bacteria, incubate these at the temperature where I want the isolated bacteria to be active (e.g. 40 °C, if using agar plates you should not exceed a temp of 50 °C), isolate single colonies and then use standard microscopic (gram staining, spore formation,....), biologic (motility, OF), and physiological (C-utilisation....) or DNA sequences to identify the genera.
Then you can start to look for your DNA ligase from the pure cultures.