BIOL 2030 Lecture Notes - Lecture 1: Restriction Enzyme, Molecular Mass, Electric Field

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salmonrat522

12 Apr 2016

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Nucleoides are negaively charged at neutral ph (because of the phosphate group) Will move towards the posiive electrode in an electric ield. Agarose is part of the agar of seaweed and uncharged. If you heat it, it will go into soluion and when they are cooled down and let to sit they will form a matrix full of pores. The comb that is inserted while the agarose is seing will be the wells. The electric current is turned on and the dna will migrate to the posiive end. Because of the pores in the agar gel the dna will be sorted according to size. Ater gel is done running you will stain gel and then expose to uv light to see where the bands are in the gel. Will migrate according to the log of their molecular mass or number of base pairs. Smaller size molecules will travel further faster than larger dna molecules.

Related Questions

1.) A restriction recognition site is repeated 1 times on a circular DNA molecule. If you treat this DNA molecule with this restriction enzyme, how many DNA fragments will you get?

a. 1

b. 2

c. 3

d. Restriction enzyme does not cut a circular molecule

2.) In an agarose gel DNA electrophoresis, the DNA fragments are separated based on their

a. Size

b. Electrical charge

c. Nucleotides sequences

d. Density

e. All of the above

3.) DNA molecules are usually separated using agarose gel rather than polyacrylamide because:

a. Agarose polymerizes faster than acrylamide gel

b. Polyacrylamide gel has pore size larger than the size of DNA molecules

c. Agarose gel do not have pores on tis matrix and can hold large DNA molecules

d. Agarose gel has larger pore size similar to the size of DNA molecules

4.) Restriction endonucleases

a. Randomly recognize and cut DNA sequences

b. Hydrolysis the phosphodiester bond of DNA backbone

c. Recognize specific sequence on double stranded DNA

d. A and B

e. B and C

taupefrog573

1. True or False: DNA fragments will "run" toward the electrode during gel electrophoresis because the groups make it a negatively charged molecule.

2. The statement that DNA fragment size is inversely proportional to mobility means: (choose the single best response)

A. That fragments above a certain size are too large to migrate through the agarose gel.

B. That polar biomolecules are best suited for this technique

C. That small fragments will migrate further than large fragments over a given time because they are able to weave through the agarose more easily.

D. None of the above are true.

3. True or False: The buffer added to the gel box during electrophoresis contains TBE to aid in conduction of electrical current.

1. On an electrophoresis power source the anode (positive charge) is red and cathode (negative charge) is black. When you put your gel in the rig, do you want the end with the comb/wells to be at the red or black end? Why?

2. The source of drag on a linear DNA molecule is the:

a. TAE Buffer

b. Nitrogen bases in the DNA molecule

c. Electric Field

d. Agarose

3. During agarose gel electrophoresis, the cause of drag on a linear DNA molecule is the:

a. Interaction with TAE buffer

b. Nitrogen bases in the DNA molecule

c. Interaction with Ethidium Bromide in the gel

d. Agarose in the gel

4. As part of your prelab work, you should have looked at the DNA isolation protocol. Why do you boil your tubes?

a. To disrupt the cell and nuclear envelope to release the DNA

b. To seperate the DNA from other cellular debris

c. To denature the DNA for PCR, the next step

d. To isolate cheek cells

5. At the end of your DNA isolation procedure you will have Chelex beads and some liquid. Explain which layer has your DNA (and thus you will save) and why.

6. What is the electric charge (q) on a 200bp linear piece of DNA?

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BIOL 2030 Lecture Notes - Lecture 1: Restriction Enzyme, Molecular Mass, Electric Field

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