SEED/2010/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 two DNA samples, you wi...) |
(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 two DNA samples, you wi...) |
(No difference)
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Latest revision as of 10:36, 2 February 2010
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 two DNA samples, you will set up 4 restriction digests using combinations of the enzymes EcoRI and PstI:
- 20ul reaction:
- 2ul NEB2
- 0.2ul BSA
- 2ul DNA (~500ng)
- 16ul DI water
- 0.5ul per enzyme
- 4 reactions
- no enzyme
- EcoRI only
- PstI only
- EcoRI and PstI
- Incubate at 37C over lunch
Prepare and pour a 1% agarose gel with 10-well combs.
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
- look at plates from [[../Day 1/]] and take quiz
- Image the gel and analyze bands
Lecture
- BioBricks & Registry of Standard Biological Parts
- DNA / restriction enzymes
- iGEM eau d'e colidemo , UT Photography
- Central Dogma Pardigm
- DNA->RNA->protein (an enzyme that makes scent)
- promoters (transcription), RBS (translation)
Instructor Preparation
- Prepare minipreps of several different BioBrick constructs in the same plasmid but with noticeably different insert lengths
- Grow up batch culture of registry:Part:BBa_J45200 with 10mM isoamyl alcohol