Difference between revisions of "Gel Extraction from Agarose"
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== Protocol == | == Protocol == | ||
| + | -Excise the DNA fragment from the gel using a scalpel and weigh it | ||
| + | |||
| + | -Add 3 volumes of Buffer QG to 1 volume of gel (100 mg~100 microliters) | ||
| + | |||
| + | -Incubate at 50 C for 10 minutes or until the gel is dissolved, vortexing every 2-3 minutes | ||
| + | |||
| + | -If the color of the mixture is violet or orange, add 10 microliters of 3M sodium acetate to lower the pH | ||
| + | |||
| + | -Add 1 gel volume of isopropanol to the sample and mix | ||
| + | |||
| + | -Put the sample into a QIAquick spin column that is in a 2 mL collection tube and centrifuge for 1 minute at 13,850 rpm. This binds the DNA | ||
| + | |||
| + | -Discard the flow through, add 0.5 mL QG Buffer and centrifuge for 1 minute at 13,850 rpm | ||
| + | |||
| + | -Discard the flow through, add 0.75 mL PE Buffer and centrifuge for 1 minute at 13,850 rpm | ||
| + | |||
| + | -Discard the flow through and centrifuge for 1 minute at 13,850 rpm | ||
| + | |||
| + | -Place the column into a fresh microfuge tube, add 50 microliters of distilled water and centrifuge for 1 minute at 13,850 rpm | ||
| + | |||
| + | -The DNA from the gel will be in the water that is in the microfuge tube | ||
== Notes == | == Notes == | ||
Revision as of 14:12, 9 July 2015
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Contents
Purpose
This protocol is used for the extraction of DNA from electrophoresis run on agarose gel.
Materials Needed
Protocol
-Excise the DNA fragment from the gel using a scalpel and weigh it
-Add 3 volumes of Buffer QG to 1 volume of gel (100 mg~100 microliters)
-Incubate at 50 C for 10 minutes or until the gel is dissolved, vortexing every 2-3 minutes
-If the color of the mixture is violet or orange, add 10 microliters of 3M sodium acetate to lower the pH
-Add 1 gel volume of isopropanol to the sample and mix
-Put the sample into a QIAquick spin column that is in a 2 mL collection tube and centrifuge for 1 minute at 13,850 rpm. This binds the DNA
-Discard the flow through, add 0.5 mL QG Buffer and centrifuge for 1 minute at 13,850 rpm
-Discard the flow through, add 0.75 mL PE Buffer and centrifuge for 1 minute at 13,850 rpm
-Discard the flow through and centrifuge for 1 minute at 13,850 rpm
-Place the column into a fresh microfuge tube, add 50 microliters of distilled water and centrifuge for 1 minute at 13,850 rpm
-The DNA from the gel will be in the water that is in the microfuge tube
Notes
Papers where this or a similar method has been used
Related Ontology Terms
References
See Help:References for how to manage references in OMPwiki.
