Stanford/BIOE44:Module 1:Day1: Difference between revisions
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=Module 1: Day 1 - Plasmid Extraction and Restriction Digest= | |||
==Introduction== | |||
==Before Class== | |||
*Please read: | *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 ([http://kirschner.med.harvard.edu/files/protocols/QIAGEN_QIAprepMiniprepKit_EN.pdf QIAPrep Spin Miniprep kit]). Then we'll measure the concentration of the plasmid DNA extracted using a nanodrop spectrophotometer. | 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 ([http://kirschner.med.harvard.edu/files/protocols/QIAGEN_QIAprepMiniprepKit_EN.pdf 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 | #Spin down culture | ||
#Resuspend | #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. | We will take the plasmid we just extracted and set up a restriction digest to cut the part out of the plasmid. | ||
Revision as of 13:57, 27 March 2010
Module 1: Day 1 - Plasmid Extraction and Restriction Digest
Introduction
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.
