Team:Wash U/Protocol
From 2009.igem.org
(Difference between revisions)
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# [[Team:Wash_U/Protocol#Glycerol Stock Solution|Glycerol Stock Solution]] | # [[Team:Wash_U/Protocol#Glycerol Stock Solution|Glycerol Stock Solution]] | ||
# [[Team:Wash_U/Protocol#Designing Primers|Designing Primers]] | # [[Team:Wash_U/Protocol#Designing Primers|Designing Primers]] | ||
+ | # [[Team:Wash_U/Protocol#Colony PCR|Colony PCR]] | ||
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=='''Mini-Prep'''== | =='''Mini-Prep'''== | ||
<font size="2"> | <font size="2"> | ||
- | :DNA Mini-Prep procedure is used to extract and isolate plasmid DNA from whole bacterial cells. | + | :DNA Mini-Prep procedure is used to extract and isolate plasmid DNA from whole bacterial cells. This Miniprep kit was obtained from Sigma-Aldrich and exact solutions and formulas are unavailable (e.g. Wash Solution 1). To get exact ingredients please contact Sigma-Aldrich. |
'''Materials''' | '''Materials''' | ||
- | + | * Wash Solution 1 | |
+ | * Wash Solution 2 | ||
+ | * dI H20 | ||
+ | * Column Prep Solution | ||
+ | * Lyse Solution | ||
+ | * Elute solution | ||
'''Procedures''' | '''Procedures''' | ||
- | : | + | # Harvest Cells |
+ | # Resuspend Cells | ||
+ | # | ||
+ | # | ||
+ | # | ||
+ | # Prepare Column: | ||
+ | # Load Cleared Lysate: | ||
+ | # Wash Column with Wash Solution 1: | ||
+ | # Wash Column with Wash Solution 2 | ||
+ | # Centrifuge at 12,000RPM for 1 minute to remove excess ethanol. | ||
+ | # Elute DNA: | ||
[[Team:Wash_U/Protocol#Procedures|Back To Top]] | [[Team:Wash_U/Protocol#Procedures|Back To Top]] | ||
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<font size="2"> | <font size="2"> | ||
:We found the Registry's protocol extremely helpful for designing primers and constructed most of our primers from this template. To visit the website, please click [http://partsregistry.org/Help:Primers/Design here]. | :We found the Registry's protocol extremely helpful for designing primers and constructed most of our primers from this template. To visit the website, please click [http://partsregistry.org/Help:Primers/Design here]. | ||
+ | [[Team:Wash_U/Protocol#Procedures|Back To Top]] | ||
+ | |||
+ | |||
+ | <font size="4"> | ||
+ | =='''Colony PCR'''== | ||
+ | <font size="2"> | ||
+ | :Description | ||
+ | '''Materials''' | ||
+ | * | ||
+ | '''Procedures''' | ||
+ | # | ||
[[Team:Wash_U/Protocol#Procedures|Back To Top]] | [[Team:Wash_U/Protocol#Procedures|Back To Top]] | ||
Revision as of 16:32, 9 July 2009