Luciferase variation among replicates

Theodosios Kyriakou theodosios73 at my-deja.com
Wed Feb 16 18:03:53 EST 2000


Variation in reporter activity might be due to at least two factors I
am aware of:
1. Difference in the number of cells you have in each well you do your
transformation in.You could resolve this by doing a simple assay about
the protein content of each well and normalise your results accordingly

2. Difference in the number of cells transfected. In the literature you
will find that do co-transfection with another plasmid containing  a
different reporter gene driven by a strong viral promoter such as RSV.

Good luck


In article <20000216045729.29926.qmail at hotmail.com>,
  pcheng18 at hotmail.com ("T Cheng") wrote:
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> Hi netter,=20
>
> I am using the luciferase assay kit to measure the luciferase
activity =
> of transfected primary culture of oyster cells in 96 well plate. The =
> luminometer were used.=20
>
> However, the luciferase activity were vary among 6 replicates of the =
> same treatment. The variation of luciferase activity were vary 100
folds =
> from 0.002 light unit (background) to 0.2 among 6 replicates.=20
>
> What are the possible cuases for this variation?=20
>
> Thanks=20
>
> Philip
>
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> <STYLE></STYLE>
> </HEAD>
> <BODY bgColor=3D#ffffff>
> <DIV><FONT face=3DArial size=3D2>
> <P>Hi netter,</FONT><FONT size=3D2> </P></FONT><FONT face=3DArial =
> size=3D2>
> <P>I am using the luciferase assay kit to measure the luciferase =
> activity of=20
> transfected primary culture of oyster cells in 96 well plate. The =
> luminometer=20
> were used. </P>
> <P>However, the luciferase activity were vary among 6 replicates of
the =
> same=20
> treatment. The variation of luciferase activity were vary 100 folds
from =
> 0.002=20
> light unit (background) to 0.2 among 6 replicates.</FONT><FONT
size=3D2> =
>
> </P></FONT><FONT face=3DArial size=3D2>
> <P>What are the possible cuases for this variation?</FONT><FONT =
> size=3D2>=20
> </P></FONT><FONT face=3DArial size=3D2>
> <P>Thanks</FONT><FONT size=3D2> </P></FONT><FONT face=3DArial
size=3D2>
> <P>Philip</P></FONT></DIV></BODY></HTML>
>
> ------=_NextPart_000_0005_01BF7808.6A539FC0--
> ---
>

--

Theodosios Kyriakou
Biological Sciences,WarwickUniversity ,CV4 7AL
tel:++44-1203-522440,++44-1203-572572 ext.30164


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