BIOL207 Study Guide - Restriction Enzyme, Ecori, Electrophoresis

113 views2 pages

limehippopotamus462

4 Dec 2014

School

University of Alberta

Department

Biology (Biological Sciences)

Course

BIOL207

Professor

Cameron Harrington

For unlimited access to Study Guides, a Grade+ subscription is required.

Document Summary

It is an enzyme that cuts at a specific site within a specific sequence. Each restriction enzyme will recognize only 1 sequence and cut at that specific sequence. e. g. ecori, psti, hindiii, haeiii: below is a picture of an agarose gel. + electrode: label where the positive and negative electrodes have to be for the dna to run properly. Explain why, based on the properties of dna, the electrodes have to be arranged in this way. Dna has an overall negative charge (phosphate groups in dna backbone). For dna to migrate, it must be attracted to the positive electrode. The bands are mand up of multiple pieces of dna that all run at the same length since they are the same size. Sample 1 contains a larger piece of dna. These are intercalating agents that stick themselves in the dna double helix and fluoresce under. We use them to visualize dna bands under uv light after running the gel.

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. Below is a map of a plasmid called pBR322:

(Cannot figure out how to post the picture)

Suppose that the pBR322 plasmid is cleaved with the restriction enzyme PstI. Suppose a PstI-digested segment of mouse DNA is then ligated into the PstI site in pBR322. This recombinant DNA molecule is then used to transform E. coli cells. a. Which antibiotic should be added to the medium to select for cells that have incorporated a recombinant plasmid? b. How can you explain the presence of bacterial colonies that are resistant to both antibiotics? c. You decide to digest your recombinant DNA molecule with BamHI and EcoRI and then analyze the digested recombinant DNA molecule on an agarose gel. How many bands will you see on the gel? d. You decide to digest your recombinant DNA molecule with BamHI and PstI and then analyze the digested recombinant DNA molecule on an agarose gel. How many bands will you see on the gel? ?a. Which antibiotic should be added to the medium to select for cells that have incorporated a recombinant plasmid?

b. How can you explain the presence of bacterial colonies that are resistant to both antibiotics?

c. You decide to digest your recombinant DNA molecule with BamHI and EcoRI and then analyze the digested recombinant DNA molecule on an agarose gel. How many bands will you see on the gel?

d. You decide to digest your recombinant DNA molecule with BamHI and PstI and then analyze the digested recombinant DNA molecule on an agarose gel. How many bands will you see on the gel?

purplebuffalo308

ANSWER ALL

CELL AND MOLECULAR

1 When running DNA samples, the comb should ideally be placed?

A) in the gel near the positive electrode of the electrophoresis

B) in the middle of the gel equidistant from both electrodes

C) in the gel near the negative electrode of the electrophoresis chamber

2 How is agarose powder made to dissolve in the buffer?

A) hot plate

B) sonication

C) microwave

D) stirring

3 When the power supply is turned on, how can you immediately tell that the gel is running?

A) electrical discharge can be observed on the electrodes.

B) Bubbles form on the electrodes.

C) The DNA begins to move through the gel.

D) The gel begins to heat up.

4 Which of the following is NOT a purpose of the loading buffer used in our laboratory?

A) It helps to visualize the DNA bands after electrophoresis is complete.

B) It contains EDTA which uninhibited nucleases.

C) It is thick and viscous allowing the DNA sample to load into the well more easily.

D) It contains a dye that makes the DNA sample easier to see.

taupewildebeest51

BIOL207 Study Guide - Restriction Enzyme, Ecori, Electrophoresis

Document Summary

Get access

Related Documents

BIOL207 Lecture Notes - Lecture 26: Ribonuclease, Plasmid, Agarose Gel Electrophoresis

BIOL207 Lecture Notes - Lecture 32: Base Pair, Northern Blot, Formamide

BIOL 1208 Lecture Notes - Lecture 1: Agarose Gel Electrophoresis, Ecori, Hindiii

Related Questions