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Volume Calculation for Mixing Reagents
There are 2 dead volumes to consider when mixing reagents online that is determined by the VALENT Software:
• About 300µL is lost during the mixing process itself, which should be accounted for in the sample calculations below.
• There is a 50µL dead volume for all reagent and mixing vials due to the limitation of probe.
Below is an example of how dead volumes work for mixing reagents.
DAB Reagent = DAB Chr DAB Buffer
The dispense volume per slide is 300µL with an extra 20µL needed, (the extra 20µL volume is to compensate for lost volume
during slide application) for a total of 320µL.
DAB Reagent
Volume for 12 slides= 12 * 320µL= 3840µL
Dead volume = 50µL
Total DAB Reagent required for 12 sides = 3890µL
a� DAB Chromogen
Volume needed for 12 slides = 240µL
Dead volume = 50µL
Total volume = 290µL
b� DAB Buffer
Volume for mixing 12 slides = 3950µL
Dead volume = 50µL
Total = 4000µL
After mixing buffer and chromogen, the total volume of DAB Reagent will be 4190µL (240µL + 3950µL). During the mixing
process, 300µL is lost leaving 3890µL, (4190µL -300µL).
Summary of Contents for VALENT
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