Table 2 Restriction endonuclease digest of genomic DNA Reaction components gDNA Hpall OR Mspl . Restriction endonuclease buffer Water Details 200 ng/μl Mspl: 20 units/μl Hpall: 10 units/μl Provided as 10x stock Add to make final reaction volume 20μl Amount in final reaction 1ug 20 units 1x in final reaction Add to make final reaction volume 20μl Volume in final reaction (μl) DNA digestion 1hr, 37°C Remember that you need to set up one tube for Mspl and 1 tube for Hpall, see Figure 7

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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8 Bb Bb Bb
616033?X-Blackboard-S3-Bucket-learn-ap-southeast-2-prod-fleet01-xythos&X-Blackboard-Expiration=1682402400000&X-Blackb
12 / 26
Reserve for analysis
using agarose gel
electrophoresis (250ng)
- 100% + B
Elective Ideas
Activity 1: Digestion of Genomic DNA with Hpall or Mspl
Water
1μg genomic DNA
20 units of Mspl per ug DNA
Reaction volume 20μl
Hpall OR Mspl*
Digest 90 mins 37°C
Take 500ng of digested DNA
Sequencing adaptor final concentration
0.5 or 1μM
Ligation reaction 10 mins
Reaction volume 20μl
Take 50ng, PCR reaction
Column-based purification of PCR products
Nanodrop quantification of products
Analyse up to 200ng
using agarose gel
electrophoresis
Reaction components
gDNA
Table 2 Restriction endonuclease digest of genomic DNA
Restriction endonuclease
buffer
b
1μg genomic DNA
20 units of Hpall per pg DNA
Reaction volume 20μl
Digest 90 mins 37°C
Take 500ng of digested DNA
Sequencing adaptor final concentration
0.5 or 1μM
Ligation reaction 10 mins
Reaction volume 20μl
Figure 7. Diagram showing the setup of digestion of human DNA with Mspl or Hpall. These
reactions are set up in 0.2ml (200μl) tubes. The gDNA is extracted from human circulating white
blood cells and is provided at a concentration of 0.2mg/ml (200ng/ul).
Take 100ng, PCR reaction
Column-based purification of PCR products
Nanodrop quantification of products
Details
200 ng/μl
Mspl: 20 units/μl
Hpall: 10 units/μl
Provided as 10x stock
Add to make final
reaction volume 20μl
SAMSUNG
Analyse up to 200ng
using agarose gel
electrophoresis
H
Amount in final
reaction
1ug
20 units
1x in final reaction
Add to make final
reaction volume
20μl
Reserve for analysis
using agarose gel
electrophoresis
W
DNA digestion
1hr, 37°C
* Remember that you need to set up one tube for Mspl and 1 tube for Hpall, see Figure 7
Volume in final
reaction (μl)
12
Transcribed Image Text:8 Q Search 8 Bb Bb Bb 616033?X-Blackboard-S3-Bucket-learn-ap-southeast-2-prod-fleet01-xythos&X-Blackboard-Expiration=1682402400000&X-Blackb 12 / 26 Reserve for analysis using agarose gel electrophoresis (250ng) - 100% + B Elective Ideas Activity 1: Digestion of Genomic DNA with Hpall or Mspl Water 1μg genomic DNA 20 units of Mspl per ug DNA Reaction volume 20μl Hpall OR Mspl* Digest 90 mins 37°C Take 500ng of digested DNA Sequencing adaptor final concentration 0.5 or 1μM Ligation reaction 10 mins Reaction volume 20μl Take 50ng, PCR reaction Column-based purification of PCR products Nanodrop quantification of products Analyse up to 200ng using agarose gel electrophoresis Reaction components gDNA Table 2 Restriction endonuclease digest of genomic DNA Restriction endonuclease buffer b 1μg genomic DNA 20 units of Hpall per pg DNA Reaction volume 20μl Digest 90 mins 37°C Take 500ng of digested DNA Sequencing adaptor final concentration 0.5 or 1μM Ligation reaction 10 mins Reaction volume 20μl Figure 7. Diagram showing the setup of digestion of human DNA with Mspl or Hpall. These reactions are set up in 0.2ml (200μl) tubes. The gDNA is extracted from human circulating white blood cells and is provided at a concentration of 0.2mg/ml (200ng/ul). Take 100ng, PCR reaction Column-based purification of PCR products Nanodrop quantification of products Details 200 ng/μl Mspl: 20 units/μl Hpall: 10 units/μl Provided as 10x stock Add to make final reaction volume 20μl SAMSUNG Analyse up to 200ng using agarose gel electrophoresis H Amount in final reaction 1ug 20 units 1x in final reaction Add to make final reaction volume 20μl Reserve for analysis using agarose gel electrophoresis W DNA digestion 1hr, 37°C * Remember that you need to set up one tube for Mspl and 1 tube for Hpall, see Figure 7 Volume in final reaction (μl) 12
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