2 replies to this topic

[Hos]

Hi guys.

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

Thanks

[phage434]

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.

[gebirgsziege]

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.