Suppose you have a sample of plasmid DNA that you want to analyze by performing a restriction digest and a gel. You have 200 μl of your DNA sample, and the DNA concentration is 40 µg/ml. The uncut DNA is 10 kb long, and you expect it to be cut into fragments of 6 kb, 3 kb, and 1 kb. You want to cut enough DNA to give you 50 ng in the 1-kb band, and then load it all on the gel. Show your calculations for everything you would put into the restriction digest tube. The stock concentration of the restriction enzyme is 5 Units/μl, and one unit of enzyme cuts one µg of DNA.

Suppose you have a sample of plasmid DNA that you want to analyze by performing a restriction digest and a gel. You have 200 μl of your DNA sample, and the DNA concentration is 40 µg/ml. The uncut DNA is 10 kb long, and you expect it to be cut into fragments of 6 kb, 3 kb, and 1 kb. You want to cut enough DNA to give you 50 ng in the 1-kb band, and then load it all on the gel. Show your calculations for everything you would put into the restriction digest tube. The stock concentration of the restriction enzyme is 5 Units/μl, and one unit of enzyme cuts one µg of DNA.

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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During electrophoresis, DNA molecules can easily be separatedaccording to size because all DNA molecules have the samecharge–mass ratio and the same shape (long rod). Would youexpect RNA molecules to behave in the same manner as DNAduring electrophoresis? Why or why not?
You digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield?
You digest 4 uL of plasmid DNA that is 50 ng/uL concetration in a total volume of 20 uL. You run 10 uL of the digest on teh gel. You then do a DNA purification protocol with a Zippy prep on the remaining digested DNA. You elute the DNA in a 25 uL. A 2 uL ssample has the concentration of 2 ng/uL. What is the DNA yield? YIELD = How much dna came out of the column/how much DNA was loaded onto the columnRemember to subtract amount of DNA run on gel from total digested DNA to get hw much DNA was loaded onto the columnAmount that came out of the column equals the volume of the eluted DNA times the concentration of the purified DNA
A piece of linear DNA is 700000 base pairs in length and is digested with a restriction enzyme that recognizes a six base pairs site. How many fragments do you predict would be produced? Show all work.
If a 500 bp of DNA between the two restriction sites were deleted, how would the banding pattern on the gel differ from the one you drew in part a? (PART A WITH THE FIRST PART OF THE QUESTION IS ATTACHED)
If a 500 bp of DNA between the two restriction sites were deleted, how would the banding pattern on the gel differ from the one you drew in part a? (Part a is attached)
Indicate the size in kilobases (kb) of DNA fragment(s) that you would expect to see if you cut a circular plasmid with BamHI recognition sites are at positions 2000bp and 3000bp and the plasmid if 5500bp total?
You are studying a new plasmid, and you digest the plasmid with three restriction enzymes: Eco RI (E), HindlII (H), and Xbal (X). You digest the plasmid DNA with each of the following combinations of enzymes and observe the results on an agarose gel. You are provided a partial plasmid map as shown below to the right. E+H E+X H+x Kb +4.3 +2.8 -+2.5 -2.0 - -1.8 +1.5 -1.0 12 F0.8 +0.5 a. What is the size of this plasmid in base pairs? b. What is the distance in base pairs between E1 and H? c. What is the distance in base pairs between E1 and X? d. What is the distance in base pairs between E2 and H? e. What is the distance in base pairs between E2 and X?
You have two tubes, each with a pair of DNA fragments inside them. Tube #1 has fragments that are 500bp and 1000 bp in length. Tube #2 has fragments that are 7500bp and 8000bp in size. If you were to perform agarose gel electrophoresis and run the contents of each tube in two separate lanes on the same gel, what would you expect to see? O That the difference between the distances migrated by the two fragments in Tube #1 was much greater than the difference between the distances migrated by the two fragments in Tube #2. O That the difference between the distances migrated by the two fragments in Tube #1 was the same as difference between the distances migrated by the two fragments in Tube #2. O That the difference between the distances migrated by the two fragments in Tube #1 was much less than the difference between the distances migrated by the two fragments in Tube #2. O It is not possible to estimate what we would expect to see.
You digest a 5 kb plasmid with two enzymes that yield fragments of 4 kb and 1 kb. After running the digest on a gel, what would you predict to see in terms of the intensities of the bands?
During gel electrophoresis, DNA molecules can easily be separated according to size because all DNA molecules have the same charge-to-mass ratio and the same shape (long rod). Would you expect RNA molecules to behave in the same manner as DNA during gel electrophoresis? Why or why not?
The total (50ul) of your restriction digest contains 500ng of DNA how much DNA (ng) are you loading into the gel?