Why do we perform Agarose Gel Electrophoresis? Prepare 4M of Magnesium Chloride

Question

Why do we perform Agarose Gel Electrophoresis? Prepare 4M of Magnesium Chloride (MW: 95.211) solution in 1 liter of water. How much volume of Stock Solution will you use to make three separate 100 ml, of 10.0 milli molar solutions from the stock solution 1.0 Motor solutions? Show your Work On your left is the picture of an agarose gel electrophoresis with six lanes. Please identify the following Which lane(s) contain a neutral molecule(s)? Which lanes contain a negative molecule(s)? Which lanes contain a positive molecule(s)? Explain what could be the possible reasons for lanes 5 and 6, which contain molecules that have similar molecular weight. What is the difference between lanes 1 and 4.

Explanation / Answer

Ans for Q.8: Agarose Gel electrophoresis:

It is a method of gel electrophoresis which is used in molecular biology and biochemistry. This technology helps to separate mixture of macromolecules such as nucleic acid and proteins in an agarose matrix. Bimolecules are separated when we applied electric field, due to this electric field charged molecules move through agarose matrix and separated by size in the agarose matrix. DNA and RNA (Nucleic acids) fragments are separated according to their length and proteins are separated according to their charge or size.

Ans for Q.9: Molarity:

Molarity defines as the number of moles of the solute in per liter of solution:

Molarity = Moles of solutes/Liters of solution

Moles= weight/Molecular weight

New formula for molarity will be, Molarity = Wt/Mol. Wt/ Liter of solution

Molecular wt of Mg Cl2= 95.21 (It is given in the question)

For the preparation of 4M solution of Mg Cl2 we have to calculate the required wt of Mg Cl2:

Wt = Molarity × Mol. Wt × Liter of solution

Wt= 4 × 95.21 × 1

Wt = 380.84 gm

When we dissolve 380.84 gm of MgCl2 in 1 L of water, it will give you 4M solution.

Ans for Q.10:

We can use the formula: M1V1= M2 V2

1M= 1000mM

M1= 1Mor 1000mM, V2=?

M2=10mM, V2= 100ml

1000 × V2= 10 × 100

V2= 1ml

For the preparation of 100ml of 10mM solution, we have to dilute 1ml of 1M stock solution up to 100ml.

So for the preparation of three separate100ml solutions we need 3ml of 1M stock solution.

Ans for Q.11:

In electrophoresis, we know that negative charge molecules attracts towards positive and positive charge molecules attract towards negative and neutral molecule will be in middle of the medium.

a.Third lane contain neutral molecule

b.1, 4, 5 and 6 lanes contain negative molecules

c.2nd lane contain positive molecules

d.Lanes 5 and 6 contain molecules of similar weight because their chrge/size ratio is also similar

e. Lanes 1contain high molecular wt molecule and 4 contain low molecular wt molecules.

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