EPPING, LQB381

Flow Diagrams

Dr Ron EppingLQB381 Unit Coordinator

1991 - 2012

EPPING, LQB381

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EPPING, LQB381

Simple diagrams help you organise your mind and understand how dilution factors apply to an experiment. Try working backwards using AMOUNTS (g or moles) from the results of the test sample to calculate the AMOUNT in the original sample.

Here is an example

100 uL of a protein extract is diluted 1/20 with saline and then assayed for protein as follows:

250 μL of the test sample (diluted extract) is mixed with 100 μL of Reagent A and 500 μL of Reagent B. The solution is then diluted to 3 mL with d.water and the absorbance measured. From comparison with plotted standards on a standard curve. the 3 mL sample is found to contain 1.5 mg of protein.

What is the protein concentration of the original solution in mg/mL?

EPPING, LQB381

Flow Diagram: Example

ProteinSolutionPROTEIN

SOLUTION

1.0 mL

EPPING, LQB381

Flow Diagram: Example

ProteinSolution

0.1mL

PROTEINSOLUTION

0.1 mL Original Sample

1.0 mL

EPPING, LQB381

Flow Diagram: Example

ProteinSolution

0.1mL

PROTEINSOLUTION

0.1 mL Original Sample

1.9 mL saline

1.0 mL

EPPING, LQB381

Flow Diagram: Example

ProteinSolution

0.1mL 0.25 mL

PROTEINSOLUTION

0.1 mL Original Sample

1.9 mL saline

1.0 mL 2.0 mL

EPPING, LQB381

Flow Diagram: Example

ProteinSolution

0.1mL 0.25 mL

PROTEINSOLUTION

0.1 mL Original Sample

1.9 mL saline0.25 mL Test Sample

1.0 mL 2.0 mL

EPPING, LQB381

Flow Diagram: Example

ProteinSolution

0.1mL 0.25 mL

PROTEINSOLUTION

0.1 mL Original Sample

1.9 mL saline0.25 mL Test Sample

1.0 mL 2.0 mL 0.25 mL

0.25 mL

EPPING, LQB381

Flow Diagram: Example

ProteinSolution

0.1mL 0.25 mL

PROTEINSOLUTION

0.1 mL Original Sample

1.9 mL saline0.25 mL Test Sample

1.0 mL 2.0 mL 0.25 mL

ASSAY PROCEDURE

+ 0.1 mL A+ 0.5 mL B+ 0.15 mL H2O

1.0 mL

ABSORBANCE

0.25 mL

1.5 mgRead from thestandard curve

EPPING, LQB381

Flow Diagram: Example

0.1mL 0.25 mL

PROTEINSOLUTION

0.1 mL Original Sample

1.9 mL saline0.25 mL Test Sample

1.0 mL 2.0 mL 0.25 mL

0.25 mL

CALCULATE USING AMOUNTS:1.5 x 10 -3 g x 2.0 / 0.25 x 1.0 / 0.1= 0.12 g in 1 mL original sample= 120 mg/mL

ASSAY PROCEDURE

+ 0.1 mL A+ 0.5 mL B+ 0.15 mL H2O

1.0 mL

ABSORBANCE

1.5 mg

EPPING, LQB381