2 replies to this topic

[Gradstudent78]

Does the way you lay your plate out matter?  Should I always follow the kit instruction manual for plate layout or doesn't matter as long as I'm consistent accross my own runs?  Is there an arguement for using one layout versus another one?  Thanks.

Grad.

Edited by Gradstudent78, 21 March 2011 - 03:04 PM.

[PAO_ahac]

Layout does not matter as long as you are consistent.  In some cases protocols demand complete randomization across the plate.  Only things to watch for are end of run effects (if assay incubation time is short and sample number is large) or contamination between very high concentration samples and low (due to poor wash technique).  Mostly layouts are standards/calibrators low to high then samples/controls.

[Gradstudent78]

PAO_ahac, on 23 March 2011 - 02:29 AM, said:

Layout does not matter as long as you are consistent.  In some cases protocols demand complete randomization across the plate.  Only things to watch for are end of run effects (if assay incubation time is short and sample number is large) or contamination between very high concentration samples and low (due to poor wash technique).  Mostly layouts are standards/calibrators low to high then samples/controls.

Thanks, could clarify what you mean by "end-of-run effects" and what can be done about it?  Is it variation accross the plate based upon when the sample is loaded (samples at the beginning of the plate different then those at the end)? I may be getting some of that.