DNA isolation from semen

[Manal Melan]

welcome


can any one here help me please.i wanna some webs talk about

DNA isolation taken from spirms " methods to isolation "

is it differ from other type of sample like DNA IN HAIR

[mvzarvg]

Hello,
The are several methods for DNA extraction from semen samples, it depends what do you want to do. I have been working with bovine semen samples the last months, and I can tell you that semen is the worst kind of cells to isolate DNA, because of the chromatin packaging by protamines.I tried all kind of protocols (solvent and no solvent). The best protocol for me is the phenol chloroform, but recently I tried Chelex 100, this resin purifies the DNA and is a good option if you only want to see if the target genes are present or not in your samples, if you need to quantify the DNA concnetration the Chelex is not a good option.

[daytwa]

How do you deal with polysaccharide contamination during your phenol/chloroform prep?

Perhaps I'm using too many cells? Or just skipping some necessary step?

Thanks

[Manal Melan]

thanks alot 4 ur help.
i needed for search only

[mvzarvg]

Hi daytwa,
I m not so sure about how you can identify polysaccharide contamination, I assume that may be the polysacharides could affect the ratios 260/280 or 260/230. In my case the big problem is the disruption of the protamines bonds to release the DNA in the sperm. I am using DTT and proteinase K to release the DNA content from the cells. But may be you could try to purify yours DNA samples with commercial columns for DNA purification. I tried once, but in my case it did not work, because the DNA concentration of my sample before purification was too low (around 15 ng/microliters). The initial concnetration of cells for my samples is around 20, 000 sperms. Good luck with your experiments.

[felipe]

Even i was not aware of the methods to isolation. Is this process takes lot of time and is there any precautions to take as done in normal DNA testing?

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