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southern BLot and agarose gel electrophoresis

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southern BLot and agarose gel electrophoresis

Postby KTK » Tue Mar 29, 2005 6:41 pm

Hi,
My question is a bit complicated...but I will really appreciate it if someone would be willing to help me out....

I'm having trouble determing from two diagrams of gels (of chromatin that was isolated from brain and liver cells and were treated with concentrations of DNaseI. The DNA is purified, cut with restriction endonuclease-- that releases the globin gene and the ovalbumin on fragments of different sizes--These fragments are then separted by agarose gel electrophoresis, transfered to nitrocelluose shees by Southern blotting and then hybridized with radiolabelledprobes specific for the globin and ovalbumin genes.) which tissue can transcribe the ovalbumin gene? In the diagrams, band 1 is identical for both the brain and the liver cells. But on band 2 the liver cell at parts 0.05 and 0.1 ug/ml are not completly fillled. What does this mean?
Thank you,
KTK
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Postby DevGrp » Thu Mar 31, 2005 9:39 am

I probably can't give a 100% accurate answer to this, just point you what I think is the right direction. This has to do with aceylation of histones in chromatin and its role in the regulation of gene expression.

The way I understand it (and please look this up) DNA of genes which can be expressed are "exposed" and sensitive to DNAse treatment. So if this follows then band 1 has been protected from the DNase and can't be expressed in either tissue. Band 2 (which I hope is ovalbumin) is sensitive in the liver so can be transcribed.

PLEASE CHECK THIS

look up
aceylation of histones in chromatin and its role in the regulation of gene expression

and senisitvity to DNAse I

Anyone else go any ideas?
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