Pin Tool Carry Over Data

     


CALCULATING CARRY-OVER BACKGROUND ON SLOT PINS AFTER 100 MOTHER/DAUGHTER TRANSFERS AND WASHES

 
 

 
VP 96 Pin Tool with FP1S50 pins (slot pin that transfers 62 nl under conditions used)

 Process steps:

Triple dip in mother plate

Triple dip in daughter plate

Blot on lint free paper

Triple dip in H2O wash reservoir (100 ml)

Blot on lint free paper

Triple dip in isopropanol wash reservoir (100 ml)

Blot on lint free paper

Repeat above process 99 times

Triple dip in assay plate and read carry-over background counts

 

I.          FITC stock concentration = 25 mg/ml in DMSO

@ 62 nl transfer = 1.55 ug FITC taken up per pin per transfer

 

II.         FITC detected in assay plate after 100 transfer/wash/blot cycles

= 0.0000924 ug* in 200 ul per well

*Data from “Carry-Over Test Plates 04-10-02”

 

III.        Percent of FITC taken up per transfer detected after final wash:

                        0.0000924 ug / 1.55 ug = 0.0000596 x 100 = 0.00596%

 
         

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