14. preparing your samples. normally your dna agarose samples include: 1. dna and 2. loading dye. sometimes…

14. preparing your samples. normally your dna agarose samples include: 1. dna and 2. loading dye. sometimes you will also add water. loading dye has multiple purposes: 1. it will help your samples sink to the bottom of the well and 2. it will help you watch the migration of your dna samples. you will be given a mock dna sample (2.5 μg/μl) and 6x loading dye. you will need to determine the appropriate total volume you will need to load with still being able to fit it in your well. common volumes are ~15 - 20 μl. prelab question - 19 (1 pt). if you want to load a total of 10 μg of dna list the volume of your dna solution you need. prelab question - 20 (1 pt). determine the total volume you will load on your gel. prelab question - 21 (1 pt). determine the volume of 6x loading dye. determine the volume of water you will need to add.

14. preparing your samples. normally your dna agarose samples include: 1. dna and 2. loading dye. sometimes you will also add water. loading dye has multiple purposes: 1. it will help your samples sink to the bottom of the well and 2. it will help you watch the migration of your dna samples. you will be given a mock dna sample (2.5 μg/μl) and 6x loading dye. you will need to determine the appropriate total volume you will need to load with still being able to fit it in your well. common volumes are ~15 - 20 μl. prelab question - 19 (1 pt). if you want to load a total of 10 μg of dna list the volume of your dna solution you need. prelab question - 20 (1 pt). determine the total volume you will load on your gel. prelab question - 21 (1 pt). determine the volume of 6x loading dye. determine the volume of water you will need to add.

Answer

Explanation:

Step1: Calculate the volume of DNA solution

Use the formula (V=\frac{m}{C}), where (m = 10\space\mu g) (mass of DNA) and (C=2.5\space\mu g/\mu l) (concentration of DNA solution). (V=\frac{10\space\mu g}{2.5\space\mu g/\mu l}=4\space\mu l)

Step2: Determine the total volume (assuming a common volume of (15\space\mu l))

Let the total volume (V_{total}=15\space\mu l)

Step3: Calculate the volume of 6X loading dye

For a 6X loading dye, a common practice is to have the loading dye be (\frac{1}{6}) of the total volume. But if we assume the total volume is (15\space\mu l), and we already have (4\space\mu l) of DNA. Let the volume of loading dye be (V_{dye}). Also, if we consider the ratio of loading dye to the final solution (a common ratio for 6X loading dye is that it is (\frac{1}{6}) of the non - DNA part. But a simpler way: assume the total volume (V_{total}) and use the formula for the volume of loading dye (V_{dye}). A common rule of thumb for 6X loading dye is that the volume of loading dye (V_{dye}=\frac{V_{total}-V_{DNA}}{5}) (since 6X means 1 part dye and 5 parts other). If (V_{total} = 15\space\mu l) and (V_{DNA}=4\space\mu l), then (V_{dye}=\frac{15 - 4}{5}= 2.2\space\mu l\approx2\space\mu l)

Step4: Calculate the volume of water

(V_{water}=V_{total}-V_{DNA}-V_{dye}). If (V_{total} = 15\space\mu l), (V_{DNA}=4\space\mu l) and (V_{dye} = 2\space\mu l), then (V_{water}=15-(4 + 2)=9\space\mu l)

Answer:

  • For the volume of DNA solution: (4\space\mu l)
  • For the total volume (assuming (15\space\mu l)): (15\space\mu l)
  • For the volume of 6X loading dye (assuming total volume (15\space\mu l)): (2\space\mu l)
  • For the volume of water (assuming total volume (15\space\mu l)): (9\space\mu l)