Hi everyone,
I would like to ask for the scientific community's help about colony PCR for generation confirmation. I have isolated bacteria from mice faeces and, after a first characterisation with Gram staining and catalase test, I would like to perform colony PCR with specific primers for Lactobacillus and Bifidobacterium:
Lactobacillus spp.: Lab-0159: 5' - GGA AAC AG (A/G) TGC TAA TAC CG -3'
Lab-0677: 5' - CAC CGC TAC ACA TGG AG -3'
Bifidobacterium spp.: OPA-02: 5' -TGC CGA GCT G -3'
OPA-07: 5' - AGG CGG GAA C -3'
Do you think that the colony PCR is really suitable for a genera confirmation?
Have you done this before?
Do you know a good PCR mix to do this?
Thank you very much for your time and your help!
Muriel
Hello, for lactobacilli and bifidobacteria better to use API test for species level identification. The genera level identification can be done without any PCR just using milk fermentation ability and betta galactosidase test. PCR can give you answer based on primer specificity but it only confirmation without cultural study.
On the contrary, in our hands, phenotypic tests like ApiZym or ApiChem identified species differently than 16s sequencing, or whole genome shotgun sequencing.
I want to do colony PCR in the future.