3. Ligation: Reaction Component Volume to Add Added? to Reaction Insert: hure EcoRI/Hindll-digested plUC19plasmid (20 ng/µl) 130 4 MI Need 80 ng Vector: EcoRI/Hindlll-digested PUC19 plasmid (20 ng/ul) 1.25 Ml 20ML 25 Need 25 ng OR 2oh Ligase enzyme 26M (10 x ) Ligase buffer 2MI 20 Water 11.75MI Total Volume 20 ul CIVI ezvz 4. Gel electrophoresis: Amount of DNA Ladder to add to gel (Lane 1): 51t Tmi ux 10cding 44Pe Volume of Volume of Sample Sample Total Volume Sample Loading Dye Undigested human 9DNA Lane 2 5 ul I MI Lane 3 Digested human 9DNA 5 µl IMI ウ-しし Undigested pUC19 plasmid Lane 4 5 µl IMI Digested pUC19 plasmid Lane 5 5 ul しM」 5. Measure the distance between electrode wires in the bottom of the gel box. At how many volts should you run your gel? (see p. 64 2b.) 13.5cm orait 6. Describe what you see on your gel in each lane. Be sure to include estimates of the sizes of DNA fragments in each lane. wwww 4444444 RR 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
3. Ligation: Reaction Component Volume to Add Added? to Reaction Insert: hure EcoRI/Hindll-digested plUC19plasmid (20 ng/µl) 130 4 MI Need 80 ng Vector: EcoRI/Hindlll-digested PUC19 plasmid (20 ng/ul) 1.25 Ml 20ML 25 Need 25 ng OR 2oh Ligase enzyme 26M (10 x ) Ligase buffer 2MI 20 Water 11.75MI Total Volume 20 ul CIVI ezvz 4. Gel electrophoresis: Amount of DNA Ladder to add to gel (Lane 1): 51t Tmi ux 10cding 44Pe Volume of Volume of Sample Sample Total Volume Sample Loading Dye Undigested human 9DNA Lane 2 5 ul I MI Lane 3 Digested human 9DNA 5 µl IMI ウ-しし Undigested pUC19 plasmid Lane 4 5 µl IMI Digested pUC19 plasmid Lane 5 5 ul しM」 5. Measure the distance between electrode wires in the bottom of the gel box. At how many volts should you run your gel? (see p. 64 2b.) 13.5cm orait 6. Describe what you see on your gel in each lane. Be sure to include estimates of the sizes of DNA fragments in each lane. wwww 4444444 RR 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
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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Transcribed Image Text:3.
Ligation:
Reaction Component
Volume to Add
Added?
to Reaction
Insert:
hure
EcoRI/Hindll-digested plUC19plasmid (20 ng/µl)
130
4 MI
Need 80 ng
Vector:
EcoRI/Hindlll-digested PUC19 plasmid (20 ng/ul)
1.25 Ml
20ML
25
Need 25 ng
OR
2oh
Ligase enzyme
26M
(10 x ) Ligase buffer
2MI
20
Water
11.75MI
Total Volume
20 ul
CIVI ezvz
4.
Gel electrophoresis: Amount of DNA Ladder to add to gel (Lane 1): 51t Tmi ux 10cding
44Pe
Volume of
Volume of
Sample
Sample
Total Volume
Sample
Loading Dye
Undigested human
9DNA
Lane 2
5 ul
I MI
Lane 3
Digested human 9DNA
5 µl
IMI
ウ-しし
Undigested pUC19
plasmid
Lane 4
5 µl
IMI
Digested pUC19
plasmid
Lane 5
5 ul
しM」
5. Measure the distance between electrode wires in the bottom of the gel box. At how many volts
should you run your gel? (see p. 64 2b.)
13.5cm orait
6. Describe what you see on your gel in each lane. Be sure to include estimates of the sizes of DNA
fragments in each lane.
wwww
4444444 RR
Transcribed Image Text: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
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