7. Did the digestion of your human 9DNA work? How can you tell? 8. Did the digestion of your PUC19 plasmid work? How can you tell? 9. What size is the digested PUC19 plasmid, according to your gel? To estimate size, simply interpolate based on the two closest bands of the DNA ladder (your lab instructor will give you the known band sizes for this). 10. Does the gel support the average predicted insert size you calculated in Lab 5 (~1600 bp)? Explain your answer. 11. Go back to the table in #3. Suppose you forgot to add the digested human gDNA to the ligation reaction. a. Would you have any ligation taking place? Explain. b. What would this look like if you ran the ligation reaction on a gel? Laboratory 6 Lane 2 Lane 3 Lane 4 Lane 5 base pairs 10000 8000 6000 5000 4000 3500 3000 Lane 1 DNA Ladder Lane 2 Uncut human gDNA (slightly degraded) Lane 3 EcoRI/Hindl-digested human 8DNA 2500 2000 Lane 4 Uncut pUC19 vector 1500 Lane 5 EcoRI/Hindll-digested pUC19 vector 1000 750 500 What is this band? 250 1% agarose gel with GelRed Lane 1

7. Did the digestion of your human 9DNA work? How can you tell? 8. Did the digestion of your PUC19 plasmid work? How can you tell? 9. What size is the digested PUC19 plasmid, according to your gel? To estimate size, simply interpolate based on the two closest bands of the DNA ladder (your lab instructor will give you the known band sizes for this). 10. Does the gel support the average predicted insert size you calculated in Lab 5 (~1600 bp)? Explain your answer. 11. Go back to the table in #3. Suppose you forgot to add the digested human gDNA to the ligation reaction. a. Would you have any ligation taking place? Explain. b. What would this look like if you ran the ligation reaction on a gel? Laboratory 6 Lane 2 Lane 3 Lane 4 Lane 5 base pairs 10000 8000 6000 5000 4000 3500 3000 Lane 1 DNA Ladder Lane 2 Uncut human gDNA (slightly degraded) Lane 3 EcoRI/Hindl-digested human 8DNA 2500 2000 Lane 4 Uncut pUC19 vector 1500 Lane 5 EcoRI/Hindll-digested pUC19 vector 1000 750 500 What is this band? 250 1% agarose gel with GelRed Lane 1

Biology: The Dynamic Science (MindTap Course List)

4th Edition

ISBN:9781305389892

Author:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Publisher:Peter J. Russell, Paul E. Hertz, Beverly McMillan

Chapter18: Dna Technologies: Making And Using Genetically Altered Organisms, And Other Applications

Section: Chapter Questions

Problem 10TYK

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1. Which of the following actions in the DNA extraction of strawberry separates the DNA from the rest of the solution? a. Addition of cold isopropyl alcohol b. crushing the strawberry c. Addition of soapy salt solution d. Filtration of the crushed strawberries 2. What does the salt in the soapy salt solution do in the DNA extraction? a. It dissolves the DNA. b. It displaced a lot of proteins to free the DNA. c. It disrupts the phospholipid bilayer. d. It precipitates the DNA.
2b) If you denatured the random 1000 bp fragments of potoroo DNA by heating them to 95°C. and then cooled them down to 60°C and allowed them to reanneal, you would find that approximately 20% of the DNA fragments renatured very rapidly, another 30% of the DNAfragments renatured moderately rapidly, and the remaining DNA fragments renatured relatively slowly. From these results, whatfraction of the potoroo genome is composed of highly repetitiveDNA? helpful Information
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1. How does PFGE separate larger fragments more efficiently than standard electrophoresis? 2. Why is SYBR green less toxic than EtBr? 3. What are the similarities and differences between Manual and Automated Sanger Sequencing? 4. What is the relationship between DNA fragment length and the distance it will run in a gel? (Restriction Enzyme Digestion)
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