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Southern blots

southern blot

A technique used for searching for a specific dNA fragment. The process is as follows:

1. separate DNA fragments by gel electrophoresis

2. Change pH of gel to basic, thus allowing disruption of H-bonds

3. blot gel with nitrocellulose paper

4. Heat paper so as to fix dNA fragments

5. probe with labelled messenger RNA or cDNA

6. wash

complementary mRNA/cDNA fragments will have hybridised.


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PCR

... fragments sizes of 3.7, 6.3, 7.0, 8.8 and 9.6 kb. Autoradiograph from southern hybridization showed that fragments 7.0 and 9.6 are radioactive, ... HindIII sites. b) A radio-labeled gene x probe is added to the Southern blots that contain the DNA. Resulting autoradiograph showed hybridisation ...

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by Missi
Wed Nov 04, 2009 5:51 am
 
Forum: Molecular Biology
Topic: PCR
Replies: 2
Views: 945

Re: Restricion map.

... probe prepared by labeling the original gene is used to analyze southern blots of three different restriction digest.prepare a map of the insert region containing the ...

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by khushi
Sat Oct 25, 2008 3:03 am
 
Forum: Molecular Biology
Topic: Restricion map.
Replies: 3
Views: 1620

Southern Blotting and Northern blottin

... are both methods for looking at sizes of particular polynucleotides. Southern blots are for DNA while Northern blots are for RNA. The methods are basically the same, though ...

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by blcr11
Thu Sep 27, 2007 3:57 pm
 
Forum: Molecular Biology
Topic: Southern Blotting and Northern blottin
Replies: 2
Views: 1625

Precaution steps for casting a gel.

... to avoid getting polymerizing regions against the glass. I was doing Southern blots; I don't know how important it is for normal check gels and whatnot, but when casting, ...

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by icequeen11
Fri Jun 15, 2007 6:53 pm
 
Forum: Molecular Biology
Topic: Precaution steps for casting a gel.
Replies: 4
Views: 4128

*** DNA and Southern Blot ***

... and so on) you can analyse the size of the fragments of the separate blots and combine this to get the restriction map. First you only use restriction enzyme 1 and do a gel electrophoresis and Southern blot and analyse the length of the fragments (complete digest) Then repeat ...

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by sdekivit
Thu Aug 10, 2006 9:40 pm
 
Forum: Genetics
Topic: *** DNA and Southern Blot ***
Replies: 11
Views: 4063


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