SEED/2008/Day 2: Difference between revisions
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(New page: == Morning == The purpose of this exercise is to learn to run a restriction digest and an agarose gel to obtain information about a DNA construct. For each of the provided tubes, you wil...) |
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Revision as of 20:19, 7 February 2008
Morning
The purpose of this exercise is to learn to run a restriction digest and an agarose gel to obtain information about a DNA construct.
For each of the provided tubes, you will set up a restriction digest using the enzymes EcoRI and PstI
- 20ul reaction:
- 2ul NEB2
- 0.2ul BSA
- 500ng DNA
- rest DI water
- 1ul EcoRI
- 1ul PstI
- Incubate at 37C over lunch
Prepare and pour a 1% agarose gel.
Afternoon
- Run a gel
- Cover gel with buffer
- Remove gel combs
- Add gel loading buffer to your restriction digests
- Carefully load samples
- Load the DNA ladder
- bacterial photography
- look at plates from [[../Day 1/]] and analyze
- Image the gel
Lecture
- DNA structure
- central dogma
- restriction enzymes
- BioBricks
- Cloning process overview
Instructor Preparation
- Prepare minipreps of several different BioBrick constructs in the same plasmid but with noticeably different insert lengths
- During lunch: copy pictures to transparencies
Instructor Post-lab
- Incubate photography plates
