Ex05_S11-Analysis-1 - Group Members: Maria Bosquez, Parxann...

Info iconThis preview shows pages 1–3. Sign up to view the full content.

View Full Document Right Arrow Icon
Group Members: Maria Bosquez, Parxann Counts, Valerie Lopez, Natalie Myers Exercise 4b & 5 Date: 2/17/11 LI Christina Hazard Unique 48835 Day: TH Room: PAI 1.18 Lab Start Time 9:00 BIO206L Spring 2011 Q.# 1-10 1 point each Total 10 points Exercise 5 Analysis To be completed as a group and turned in at the beginning of your next laboratory period. Include your “Data & Results”, sketches, acquired digital images, etc. as directed by your laboratory instructor. Show your work for all calculations and/or print your MS Excel o data sheets. Be sure to include proper units where necessary. Adhere to University’s Honor Code and course policies. Balance between brevity and completeness. 1. Demonstrate your understanding of agarose gel electrophoresis. a. How does agarose gel electrophoresis work to separate different size fragments of DNA? DNA is negatively charged. When put into the agarose mix and then into the electrophoresis apparatus, which is positively charged at the anode end and negatively charged at the cathode end, the DNA migrates toward the positive side due to its attraction. b. What are the functions of each of the components of the “6 X DNA Loading dye” solution? The 6X DNA Loading Dye allows the DNA to be visible as it migrates across the agarose mix toward the anode side of the apparatus. c. If one wanted to separate DNA fragments of very large sizes, would one increase or decrease the percentage of the agarose in solution to make the gel? For example, which would be better: a 0.7% or 2% agarose gel to separate kb fragments? Explain your answer. The agarose gel allows the DNA fragments to migrate through its pores. Smaller fragments are able to migrate faster and farther in the agarose through its pores. By decreasing the percentage in agarose solution the longer fragments would be able to migrate even if they cannot be transported by the pores. d. If you wanted your gel to “run faster” would you decrease the ionic strength of the TAE running buffer (make it more dilute) or use more concentrated TAE running buffer (increase the ionic strength)? Explain your answer. Although the TAE running buffer allows for better resolution of DNA, its use requires low voltage which takes more time. If one wanted to run their experiment in a shorter amount of time it would be to their advantage to decrease the ionic strength. e. List some other factors that would affect the rate of DNA migration through the gel? Length, shape (conformation) of the DNA molecule and plasmid, concentration of the agarose gel, voltage 2. Prepare a standard curve showing the relationship between known size (in bp) of the molecular weight marker/ fragments (bands of the MW ladder) and the migration distance for each of the bands.
Background image of page 1

Info iconThis preview has intentionally blurred sections. Sign up to view the full version.

View Full DocumentRight Arrow Icon
Group Members: Maria Bosquez, Parxann Counts, Valerie Lopez, Natalie Myers Exercise 4b & 5 Date: 2/17/11 LI Christina Hazard Unique 48835 Day: TH Room: PAI 1.18 Lab Start Time 9:00 a. Use data from the agarose gel run to prepare a scatter plot in MS Excel. Place the
Background image of page 2
Image of page 3
This is the end of the preview. Sign up to access the rest of the document.

Page1 / 11

Ex05_S11-Analysis-1 - Group Members: Maria Bosquez, Parxann...

This preview shows document pages 1 - 3. Sign up to view the full document.

View Full Document Right Arrow Icon
Ask a homework question - tutors are online