Analys of Genomic DNA by Southern Hybridization (Southern Blot)
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Analys of Genomic DNA by Southern Hybridization (Southern Blot)
From Heiner Müller lab.
Localization of particular sequences within genomic DNA is usually accomplished by the transfer techniques described by Southern (1975). Genomic DNA is digested with one or more restriction enzymes, and the resulting fragments are separated according to size by electrophoresis through an agarose gel. The DNA is then denatured in situ and transferred from the gel to a solid support (usually nitrocellulose filter or nylon membrane). The relative positions of the DNA fragments are preserved during their transfer to the filter. The DNA attached to the filter is hybridized to radiolabeled DNA or RNA, and autoradiography is used to locate the positions of bands complementary to the probe. A sequence of 1000 bp that occurs only once in the mammalian genome can be detected in an overnight exposure if 10 µg of genomic DNA is transferred to the filter and hybridized to a probe several hundred nucleotides in length.
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Last update 16-Sep-2005, Rating Good of 4 votes.
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Please, does anyone know which side of the gel should face up or come in contact with the membrane during Southern blot transfer.
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I agree with Piyu. The information is really very educative. I am sure students like me will certainly benefit. I'll request Piyu to share information with me regarding this.
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This is realy very educative.This will defenately help students in there studies.
I request u to please send me more matter regarding this like theory,pictures/photographs if possible.
THANX
Piyu Sukhwal
3-e Shive Park Durga Nursery Road
Udaipur 313001 (Rajasthan)
INDIA
bitto_sukhwal@yahoo.com
chamcham_hi@yahoo.com
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Southern Hybridization is a very good technique but nowadays exists DNA Chip that is is a revolutionary new tool used to identify mutations in genes like BRCA1 and BRCA2. The chip, which consists of a small glass plate encased in plastic, is manufactured somewhat like a computer microchip. On the surface, each chip contains thousands of short, synthetic, single-stranded DNA sequences, which together, add up to the normal gene in question.
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