I am carrying out fluorescent in situ on the bacteria and fungi found in the GI tract of a eukaryotic organism. The bacteria is easy, EUB338 is a universal bacteria probe However the fungi appears more tricky, from what I have read EUK516 is a universal eukaryotic probe but that may also fluoresce the organisms cells too. I know there is a PF2 that only does yeast, are there any other just fungal probes?
You may use Panfungal PCR and multiplex liquid hybridization technique. For detailed procedure please consult Journal of Clinical Microbiology (2000), 38 (11), 4186-4192.
Thank you
This procedure wouldn't be of use to me. I have done Illumina sequencing (bacteria and fungal) on the organisms guts, I now need to confirm the location of the bacteria and fungi to make sure they are within the gut lumen and not on the outside of the gut. So I will be slicing the guts onto slide and doing in situ on those slices. I will be using EUB338 for bacteria but need probe for fungi.
Any ideas?
Yes I am going to is a fluorescently labelled probe but I am concerned that the probe will also target the eukaryotic organism and the fungi, not just the fungi. So I am asking if anyone knows of probes that are fungal specific, not just eukaryotic cells
You may try with oligonucleotide probes (ITS2 rDNA sequences) or cloned DNA probes, they are fungal specific.
Are they usable in FISH? I thought they were only used in pcr, I’ve not seen any papers using ITS rDNA primers in FISH, only rRNA primers
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