Southern blotting: Difference between revisions
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Southern blotting is when you isolate a particular gene. | Southern blotting is when you isolate a particular [[Gene|gene]]. | ||
First you have to do electrophoresis on with the DNA. You use restriction enzymes to break up the DNA and the run the gel using agarose. The smallest fragments ith travel the furthest. Place nitrocellulous filter on plate to pick up phages from each plaque. Left with single stranded DNA. Hybridise with labelled probe. Denature the DNA using NaOH. Left with single stranded DNA. Hybridise with labelled probe. Perform autoradiography. Signal appears over phage DNA that is complementary to probe. Cut out signal area to obtain DNA. | First you have to do [[Electrophoresis|electrophoresis]] on with the [[DNA|DNA]]. You use [[restriction enzyme|restriction enzymes]] to break up the DNA and the run the gel using [[Agarose|agarose]]. The smallest fragments ith travel the furthest. Place [[nitrocellulous|nitrocellulous]] filter on plate to pick up phages from each [[plaque|plaque]]. Left with single stranded DNA. [[Hybridise|Hybridise]] with [[labelled probe|labelled probe]]. Denature the DNA using [[NaOH|NaOH]]. Left with single stranded DNA. Hybridise with labelled probe. Perform [[autoradiography|autoradiography]]. Signal appears over [[phage|phage]] DNA that is complementary to probe. Cut out signal area to obtain DNA <ref>http://askabiologist.asu.edu/southern-blotting</ref>. | ||
<references /> | === References === | ||
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Revision as of 10:41, 12 November 2011
Southern blotting is when you isolate a particular gene.
First you have to do electrophoresis on with the DNA. You use restriction enzymes to break up the DNA and the run the gel using agarose. The smallest fragments ith travel the furthest. Place nitrocellulous filter on plate to pick up phages from each plaque. Left with single stranded DNA. Hybridise with labelled probe. Denature the DNA using NaOH. Left with single stranded DNA. Hybridise with labelled probe. Perform autoradiography. Signal appears over phage DNA that is complementary to probe. Cut out signal area to obtain DNA [1].