Stanford/BIOE44:Module 1:Day1: Difference between revisions
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===Part 2: Restriction Enzyme Digest=== | ===Part 2: Restriction Enzyme Digest=== | ||
We will take the plasmid we just extracted and set up a restriction digest to cut the part out of the plasmid. | We will take the plasmid we just extracted and set up a restriction digest to cut the part out of the plasmid. | ||
==For Next Time== |
Revision as of 14:01, 27 March 2010
M1: Day 1 - Plasmid Extraction and Restriction Digest
Introduction
- plasmids
- mini preps - purpose of each step
- parts
- restriction enzymes
- components of a digest
Before Class
- Please read:
In Class
Part 1: Plasmid extraction (aka miniprep)
At the beginning of class you will be given a 5mL culture of E. coli harboring a plasmid that contains a BioBrick part. We will extract the plasmid from the bacteria using a kit (QIAPrep Spin Miniprep kit). Then we'll measure the concentration of the plasmid DNA extracted using a nanodrop spectrophotometer.
Mini-Prep Protocol
- Spin down culture
- Resuspend
Part 2: Restriction Enzyme Digest
We will take the plasmid we just extracted and set up a restriction digest to cut the part out of the plasmid.