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Minimum concentration of bacteria to RNA extraction and Microarray


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#1 DrRibeiro

DrRibeiro

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Posted 13 November 2010 - 07:58 PM

Hello all,

I need a help/advice. Whats the minimum amount o bacteria/innoculum (10^6, 10^7, etc) that I should use to perform a RNA extraction (Trisol) and then carry out a microarray experiment??

Im asking because I want to analyse the gene expression during lag phase of L.monocytogenes and I just found one paper that extracted RNA from 10^5 UFC/ml but using 750ml cultures (too much) and likewise I found some papers that extracted from 10^7 and mostly 10^9...

My problem: I need to perform a growth curve (that last 15 days - salt included) and then extract the RNA from nearly 48 hours (lag phase). So, I need to choose the correct innoculum to get a good amount of RNA (cDNA - Microarray).

Thanks in advanced for any suggestion ou experience.

Vinicius




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