When adding your DNA samples to the agarose gel, you will need to add loading dy

Question

When adding your DNA samples to the agarose gel, you will need to add loading dye to the samples. Loading dye contains: ¥ glycerol, which helps the samples sink into the sample wells of the gel; and ¥ tracking dyes so that you have a way of estimating how far the samples have travelled through the gel during electrophoresis. The recipe for 6X loading dye is as follows: ¥ 0.25% bromophenol blue (w/v) ¥ 0.25% xylene cyanol (w/v) ¥ 30% glycerol in water (v/v) 11. Provide a recipe for 10 ml of 10X loading dye, using the information above. (Important tip: Remember that you have already made 30ml of a 50% stock solution of glycerol. I recommend determining the concentration of glycerol required for 10X loading dye, then figuring out how much of your 50% glycerol stock will be needed to make 10ml of 10X loading dye.)

Explanation / Answer

6X loading dye composition:

0.25% bromophenol blue (w/v)

0.25% xylene cyanol (w/v)

30% glycerol in water (v/v)

Required volume = 10 mL

Required concentration = 10X

i. 0.25% bromophenol blue (w/v)

1 g in 10 mL = 10%

0.1 g in 10 mL = 1%

0.025 g in 10 mL = 0.25%

ii. 0.25% xylene cyanol (w/v)

1 g in 10 mL = 10%

0.1 g in 10 mL = 1%

0.025 g in 10 mL = 0.25%

iii. 30% glycerol in water (v/v)

Stock = 50%

1 mL in 10 mL = 5%

6 mL in 10 mL = 30%

Take 3 mL of distilled water. Add required amounts of Bromophenol blue and xylene cyanol. Mix them completely. Then add 6 mL of 50% glycerol. Mix well. Make up the volume to 10 mL with water.

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