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? 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). 9. 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 9DNA 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? 3. Ligation: Volume to Add Reaction Component Added? to Reaction Insert: hume genomic EcoRI/Hindlll-digested pUC19 plasmid (20 ng/ul) Need 80 ng 4 MI Vector: EcoRI/Hindlll-digested pUC19 plasmid (20 ng/ul) Need 25 ng 1.25 Ml 20M 25 5aM Toan Ligase enzyme IMI 25M- (1o x) Ligase buffer 20 Water 11.75MI Total Volume 20 ul CIVI= ezvz 4. Gel electrophoresis: Amount of DNA Ladder to add to gel (Lane 1):5u1tlMl 1000ng Sample Sample Volume of Volume of Sample Loading Dye Total Volume Lane 2 Undigested human 5 ul
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? 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). 9. 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 9DNA 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? 3. Ligation: Volume to Add Reaction Component Added? to Reaction Insert: hume genomic EcoRI/Hindlll-digested pUC19 plasmid (20 ng/ul) Need 80 ng 4 MI Vector: EcoRI/Hindlll-digested pUC19 plasmid (20 ng/ul) Need 25 ng 1.25 Ml 20M 25 5aM Toan Ligase enzyme IMI 25M- (1o x) Ligase buffer 20 Water 11.75MI Total Volume 20 ul CIVI= ezvz 4. Gel electrophoresis: Amount of DNA Ladder to add to gel (Lane 1):5u1tlMl 1000ng Sample Sample Volume of Volume of Sample Loading Dye Total Volume Lane 2 Undigested human 5 ul
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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Need question 11 answered
Transcribed Image Text: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?
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).
9.
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 9DNA 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?
Transcribed Image Text:3. Ligation:
Volume to Add
Reaction Component
Added?
to Reaction
Insert:
hume genomic
EcoRI/Hindlll-digested pUC19 plasmid (20 ng/ul)
Need 80 ng
4 MI
Vector:
EcoRI/Hindlll-digested pUC19 plasmid (20 ng/ul)
Need 25 ng
1.25 Ml
20M
25
5aM
Toan
Ligase enzyme
IMI
25M-
(1o x) Ligase buffer
20
Water
11.75MI
Total Volume
20 ul
CIVI= ezvz
4. Gel electrophoresis: Amount of DNA Ladder to add to gel (Lane 1):5u1tlMl 1000ng
Sample
Sample
Volume of
Volume of
Sample
Loading Dye
Total Volume
Lane 2
Undigested human
5 ul
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