Gel Extraction from Agarose

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Purpose

This protocol is used for the extraction of DNA from electrophoresis run on agarose gel.

Materials Needed

-scalpel

-scale

-Buffer QG

-Buffer PE

-microfuge tubes

-centrifuge

-50 C incubator

-3 M sodium acetate

-QIAquick spin column

-distilled water

-isopropanol

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

References

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