Dear Colleagues,
The sample is serum and, according to our calculations, we have to make a 1:5000 dilution so that the values fall within the standard line.
The question we have is how to perform this dilution:
(1) Perform first a 1:5 dilution and then a 1:1000?
(2) First perform a 1:50 dilution followed by a 1:100 dilution?
Depending on whether we obtain the 1:5000 dilution one way or the other, the volume of initial serum varies; being 24 microliters for #1 and 3 microliters for form #2.
Thank you very much.
Hello Arkaitz Mucientes
I agree with Luis Mata
As far as possible use large volumes because they are easier to pipette accurately than small ones. Pipetting small amount (3ul for the form #2) accurately is not easy and will likely contribute to statistical error in your results. Factors that impact air pressure or pipette tip function can lead to volume errors, particularly when you work with small volumes.
On the other hand, when you work with large volume and pipette 24µl (for the form #1), you may easily and accurately pipette this amount, which would yield much tighter error bars.
Best.
Thanks a lot Luis Mata and Malcolm Nobre for your responses.
We will follow your advise and work with the largest volume at first.
Best regards,
Arkaitz.
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