Latest revision as of 16:03, 28 October 2008

Newcastle University

GOLD MEDAL WINNER 2008

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Home >> Wet Lab >> Protocols >> Cutting a Specific Band from Agarose Gel

A gel in the UV light box prior to cutting

Jess cutting out a fragment from the gel using a sterile scalpel blade

After running the restricted plasmid on agarose gel (prefereably with a wide comb to increase well size), identify the specific band to be cut out.

Weigh a DNase-free 1.5 ml microcentrifuge tube and record the weight.

Place the gel in a UV light box (N.B. wear gloves and a UV filter mask and ensure sleeves are completely covering wrists to avoid direct UV light exposure).

Using a sterilised scalpel blade, long wavelength(365 nm) and minimal exposure time, cut out the required band, including as little of the surrounding gel as possible.

Place agarose gel band into a DNase-free 1.5 ml microcentrifuge tube.

Weigh the microcentrifuge tube plus agarose band and calculate the weight of the actual agarose slice.

Proceed to Purifying DNA from Gel Slices.

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-[[Image:UVgel.jpg|thumb|right|300px|Our gel in the UV light box prior to cutting the band out]]
+{{:Team:Newcastle University/Header}}
+{{:Team:Newcastle University/Template:UnderTheProtocol|page-title=[[Cutting a Specific Band from Agarose Gel]]}}
-[[Image:jess_gel.jpg|thumb|right|250px|Jess cutting out the 2.2kb fragment from the gel using a sterile scalpel blade]]
+[[Image:UVgel.jpg|300px]]
+A gel in the UV light box prior to cutting
+[[Image:jess_gel.jpg|thumb|right|250px|Jess cutting out a fragment from the gel using a sterile scalpel blade]]
+* After running the restricted plasmid on agarose gel (prefereably with a wide comb to increase well size), identify the specific band to be cut out.
+* Weigh a DNase-free 1.5 ml microcentrifuge tube and record the weight.
+* Place the gel in a UV light box (N.B. wear gloves and a UV filter mask and ensure sleeves are completely covering wrists to avoid direct UV light exposure).
+* Using a sterilised scalpel blade, long wavelength(365 nm) and minimal exposure time, cut out the required band, including as little of the surrounding gel as possible.
+* Place agarose gel band into a DNase-free 1.5 ml microcentrifuge tube.
+* Weigh the microcentrifuge tube plus agarose band and calculate the weight of the actual agarose slice.
-* After running the restricted plasmid on an agarose gel (prefereably large-welled with as much plasmid as possible), identify the specific band to be cut out.
-* Record the weight of an Eppendorf tube.
-Place the gel on a UV light box (N.B. wear gloves and a UV filter mask and ensure sleeves are completely covering wrists to avoid direct UV light exposure).
-* Using a scalpel blade that has been cleaned with ethanol, carefully cut out the band. Try to include as little of the surrounding gel as possible.
-* Place the gel slice in the Eppendorf tube.
-* Record the weight the Eppendorf tube and gel slice and work out the difference between this and the empty tube. This information is important for determining how much buffer to use for purifying the DNA from the gel.
 * Proceed to [[Purifying DNA from Gel Slices]].