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Quantifying ingested bacteria without realtime PCR


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#1 umam

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Posted 06 July 2009 - 07:07 AM

Hi,

I am working with mosquito larvae and I feed them with suspended and biofilm treated GFP bacteria. I want to quantify (without using qRTPCR) how many of these are ingested by the larvae? Is there a good way to do this? Please help.
Thanks in advance,
-Uma

#2 fishdoc

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Posted 09 July 2009 - 06:22 AM

Hi,

I am working with mosquito larvae and I feed them with suspended and biofilm treated GFP bacteria. I want to quantify (without using qRTPCR) how many of these are ingested by the larvae? Is there a good way to do this? Please help.
Thanks in advance,
-Uma



Do the larvae need to live for further experimentation?

If not, homogenize the larvae in PBS and spread aliquots on agar plates and count the colonies.

#3 umam

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Posted 09 July 2009 - 06:25 AM

We have tried that but a lot of non specific bacteria grow up and we need to isolate just the one we are looking for. Addition of antibiotics to the plates also has not helped much.

#4 fishdoc

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Posted 09 July 2009 - 06:31 AM

We have tried that but a lot of non specific bacteria grow up and we need to isolate just the one we are looking for. Addition of antibiotics to the plates also has not helped much.



The GFP doesn't distinguish the one you're looking for from the others? Is the GFP on a plasmid or in the genome?




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