I ran a bunch of blank samples in a plate and noticed that towards the end of the plate samples are having a higher background then samples at the beginning of the plate. For example the blanks in wells A1 and A2 average ~0.058 optical density while H9 amd H10 ~0.083. What could be responsible for this? Thanks.
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PAO_ahac
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Posted 31 March 2011 - 04:27 AM
What is your %CV of replicates? Could be assay variability, carry over, manual v. automated washing, insufficient washing etc.
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Gradstudent78
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Posted 31 March 2011 - 05:44 AM
PAO_ahac, on 31 March 2011 - 04:27 AM, said:
What is your %CV of replicates? Could be assay variability, carry over, manual v. automated washing, insufficient washing etc.
%CV for replicates are all < 10%. I use an automated 4x washing.
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mdfenko
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Posted 31 March 2011 - 08:27 AM
does your reader read the whole plate at once or does it read a row at a time?
do you kill the color reaction or just read it?
if the reader does a row at a time and you don't kill the reactions (all at the same time) then the reaction continues during reading.
do you kill the color reaction or just read it?
if the reader does a row at a time and you don't kill the reactions (all at the same time) then the reaction continues during reading.
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Gradstudent78
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Posted 31 March 2011 - 09:30 AM
mdfenko, on 31 March 2011 - 08:27 AM, said:
does your reader read the whole plate at once or does it read a row at a time?
do you kill the color reaction or just read it?
if the reader does a row at a time and you don't kill the reactions (all at the same time) then the reaction continues during reading.
do you kill the color reaction or just read it?
if the reader does a row at a time and you don't kill the reactions (all at the same time) then the reaction continues during reading.
Not sure about the reader (it's fairly old, probably going to buy a new one soon), but we do use a stop solution to kill the reaction.
Edited by Gradstudent78, 31 March 2011 - 09:37 AM.
