Extracting and drying human DNA

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Johann

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Oct 14, 2014, 2:44:25 AM10/14/14
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Hello DIYbiologists,

i planed to do (qualititativ) detection reactions of the atoms, which my DNA is build of. Therefor i need lots of DNA. To extract DNA i used the saltwater-dishcleaner-alcohol-method(used spiritus instead of iso-propanol, worked better in my case). The problem is that i also get lots of bacterial DNA using this method. Do you have any method to extract human DNA with much less bacteria DNA, without hurt and provide a good amount DNA per charge and can be done multiple times in a row. Due i have support from a chemistry teacher i can access some chemicals.
I do not ask for a method to replicate the DNA after extracted with nucleoacidsand thermocycler.

Do you have an idea for DNA extracting, which woks better in bigger scale than the described method?

Thanks for inspiration, literature, ideas or protocols

Nathan McCorkle

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Oct 14, 2014, 1:29:30 PM10/14/14
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Drying can be done by washing the DNA with ethanol then warming with an open top in an oven.

If you're in a lab, and there are some cell/molbio folks around, you might try immortalizing some of your cells... but at that point why not just use an existing human cell line? There are only so many cells you can lose (from yourself) without noticing too much.

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SC

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Oct 14, 2014, 6:55:55 PM10/14/14
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Well, there's a way for males to extract a large amount of bacterial-free DNA from themselves...
As far as multiple times in a row however, I guess that would depend on the guy.
 
 

Dakota Hamill

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Oct 15, 2014, 1:13:40 AM10/15/14
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hahaha SC has the best recommendation besides bleeding yourself, and it'd probably feel better.

I don't really understand what you're trying to study though..."qualitative detection reactions of the atoms?"  I get that language is a barrier but that sentence doesn't really make any sense.

Mein Duetsch ist nicht so gut aber....we have a few native German speakers so if you posed the question in your native language you might get more detailed responses.  

Johann

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Oct 15, 2014, 2:49:34 PM10/15/14
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Due common request my question in improved english:

Hello DIY-bios,

i am new to this mail list. To become familiar with the very basics of DIYBio, i try some very easy experiments. But since walking the same path, as somebody did before becomes quickly boring. I choose not only to extract DNA, which i did successful before, i am planning to do detection reactions for carbon, nitrogen, hydrogen, oxygen and phosphor. In the first step i want to show that these elements are part of my sample, in the second step i extend my experiment to do quantitative measurements. It is clear that these experiments will not change the world nor help anyone, but my self to get more familiar with the scientific method and experimenting. I am going to need at least for the second step a reasonable amount of DNA, to become precise results. The idea from McCorkel to ask some Cell-Bios does not help me. The reason is my environment, i am going to do this experiment in the chemistry lab of my school in some sort of chemistry-club. That is also why i can not archive SC's hilarious idea, which carry risk being kicked out from the group.

Do you have an practical and appropriate idea to gain human lots of DNA in a school lab  during some sort of chemistry-club?

I hope the improved text lead to interesting and helpful.
Beside my question SC do you have experience with DNA extraction from spermatozoa? Is there some sort of protocol out there? Spermatozoa die at slight ph change, would this also disassemble the DNA and lead to wrong results?

Nathan McCorkle

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Oct 15, 2014, 4:59:59 PM10/15/14
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On Wed, Oct 15, 2014 at 11:49 AM, Johann <joht...@gmail.com> wrote:
Due common request my question in improved english:

Hello DIY-bios,

i am new to this mail list. To become familiar with the very basics of DIYBio, i try some very easy experiments. But since walking the same path, as somebody did before becomes quickly boring. I choose not only to extract DNA, which i did successful before, i am planning to do detection reactions for carbon, nitrogen, hydrogen, oxygen and phosphor. In the first step i want to show that these elements are part of my sample, in the second step i extend my experiment to do quantitative measurements. It is clear that these experiments will not change the world nor help anyone, but my self to get more familiar with the scientific method and experimenting. I am going to need at least for the second step a reasonable amount of DNA, to become precise results. The idea from McCorkel to ask some Cell-Bios does not help me. The reason is my environment, i am going to do this experiment in the chemistry lab of my school in some sort of chemistry-club. That is also why i can not archive SC's hilarious idea, which carry risk being kicked out from the group.

SC's idea was something I alluded to but didn't come out and say it... but really, what part of your humanity are you willing to give up to get this done? If cheek swab/ mouth rinse isn't sufficient, you only have the choice to cut a chunk of your flesh, bleed yourself, immortalize a cell line and grow it up, or use gametes which are dispensed in males quite easily. The issue of pH shouldn't matter, since you're interested in the DNA. Why not use a cell line? Or bacteria? Or yeast? Their DNA isn't know to be atomically different.


Does your school have an SEM with an EDX detector or a backscatter detector (and use ebsd)? Or a TEM and do electron diffraction spectroscopy. You can do chemical mapping with those techniques, with SEM you're probably limited to a few nanometers of resolution (and a double helix of DNA is only about 25nm wide), but won't likely get you single-atom data unless the microscope has a piezo stage which can move with sub-nanometer steps. The TEM would give you single-atom resolution, but in either case your data will be a bit muddled unless you straighten the DNA prior to viewing (with the water-air interface method). This paper gives that method and mentions a bit on TEM:


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