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Luciferase: too much pbs


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#1 Iluvkittens

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Posted 08 August 2012 - 07:53 PM

If I double the amount of PBS per well of cells that a luciferase protocol calls for, should I double the reporter lysis buffer as well? Do I then add twice as many cells to my plate when I set it up for reading on the luminometer?

#2 bob1

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Posted 09 August 2012 - 01:03 AM

It depends on what the roe of the PBS is? - do you remove it before adding the lysis buffer? if so, it is a wash step and you can proceed without doubling.

#3 Iluvkittens

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Posted 09 August 2012 - 04:40 AM

No, it's not removed. Lysis buffer is added directly.

If I double the amount of lysis buffer, do I also double the amount of assay and substrate I use (200ul instead of 100ul)?

Edited by Iluvkittens, 09 August 2012 - 05:21 AM.


#4 bob1

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Posted 09 August 2012 - 01:23 PM

Probably, as you will need to maintain a minimum concentration for those reagents.

#5 Iluvkittens

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Posted 09 August 2012 - 03:48 PM

thanks!




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