I am using 10 microlitre of test sample, 10 microlitre of microbial suspension (in hepes, adjusted to 1000 bacterial cells), and 80 microlitre of MH broth in a 96 well microtitre plate. Reading of my replicates vary a lot. I am very sure that its not because of pipetting error or error with microplate reader. Can anyone suggest a good solution?
Dear Naveen,
Perhaps If you used a dry incubation, evaporation in this step and edge effect on microplate can impact your results.
Best regards
Dear Naveen
What do you mean by replicates: wells on the same plate (i.e. dilution of the same cell suspension) or independent replicates (different cell suspensions, e.g. on different days)?
Jack
Did you well mix the cultures and your test samples in 96-well plates? Did you look at the condensed water on the lid when you do the OD reading?
@ jack Pronk....By replicates I meant wells on same plate having same dilution of sample and same cell suspension..
@Deng...yes cultures were mixed properly.
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