Dear all,
I have been testing some compounds in an assay by using 384-well plate. Compounds are in another plate and transferred into my assay plate (same as HTS) simultaneously. Because I have only one plate, I don't use an automated system to dispense reagents during the assay. I use a 12-channel pipette. In the end of the experiment, I get variation between replicates. But variation doesn't seem random. It looks like there is a column effect. You can find a picture of the plate map and outcome values attached under this question. Basically, there are 3 replicates side-by-side and 2 doses shown with two different colours. Has anyone experienced similar problem? Thanks.
Regards,
Burak
hi, did you adjust your 12 channel pipette to make sure there is a appropriate volume of pre-/post-dispense? this step help you minimize the variation of volume added into three replicated wells. it should be achieved easily if you use one INTEGRA pipette. BTW, if have air bubbles inside tips, change them before next aspirate.
Mert Burak Ozturk If you are talking about variation in the result/outcome, I might quote that there will always be variation between replicates, at least 1% in general, which can be reduced by careful handling, but not completely eliminated due to several factors like mix distribution, pipetting errors (which might be probable in your case) and human-errors which all are to be accounted for. You might check these following things to make sure your variation is reduced.
If you still face problems, feel free to discuss!
Are you changing tips every time you dispense one set of 12 samples? If not, the residual volume in the tip will introduce a source of variation (and you risk cross-contaminating your original samples). Tips are cheap, reagents & your time are valuable.
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