Mail DNA/Proteins to anyone

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Daniel

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Feb 25, 2009, 8:21:42 AM2/25/09
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Hi all,

Has anyone practically tried this in real time to "Send DNA/Proteins
stored on Filter Paper"
and mailed across to different destination and used???

REF:
Alternative Means of DNA Preservation: Dry Storage on Qualitative
Filter Paper
by Megan K. Morikawa
www.usc.edu/CSSF/History/2007/Projects/S0416.pdf

Waiting for all the comments

Daniel Singh

Jeswin John

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Feb 25, 2009, 8:26:05 AM2/25/09
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That's a science fair project! Wow, her work is very advanced at least compared to what I did. Reading it, I think it would work. Anyone know how to extract the DNA from the paper?
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Dan

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Feb 25, 2009, 8:28:52 AM2/25/09
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On Wed, Feb 25, 2009 at 05:21:42AM -0800, Daniel wrote:
>
> Hi all,
>
> Has anyone practically tried this in real time to "Send DNA/Proteins
> stored on Filter Paper"
> and mailed across to different destination and used???

I certainly used to get things this way when I worked in the lab - plasmids anyway. I wouldn't say it was utterly reliable. I think the iGEM biobricks were shipped similarly last year? I know our team couldn't get any of them to work.

I do remember seeing someone from RIKEN holding up a book of clones at a conference some years ago.

http://www.ncbi.nlm.nih.gov/pubmed/12819147

would appear to be the reference

regards,

Dan
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sgt york

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Feb 25, 2009, 9:24:04 AM2/25/09
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DNA yes, proteins no. I used to ship DNA this way & had DNA shipped to
me this way on a regular basis. I used to use filter paper (a thick
absorbant paper) with a spot of DNA dried onto it.

I have never heard of anyone doing this with proteins, though. Those
should be shipped on ice. On second thought, you could probably ship
things like western blots like this; but not functional proteins.

Daniel

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Feb 25, 2009, 9:27:17 AM2/25/09
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if not paper any other material which can be used for proteins and
what abt rna.

Dan

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Feb 25, 2009, 9:31:01 AM2/25/09
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On Wed, Feb 25, 2009 at 06:27:17AM -0800, Daniel wrote:
>
> if not paper any other material which can be used for proteins and
> what abt rna.

RNA is pretty tempermental stuff to work with, I doubt very much you could get away with doing that on filter paper.

Mackenzie Cowell

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Feb 25, 2009, 4:49:59 PM2/25/09
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That makes sense.  But just as RNA is a much more unstable encoding of the transcript of a gene, I wonder if there are ways to isolate different states of DNA that is even hardier than, say, what you would get with a miniprep.  For instance, perhaps in a particular stage of growth the cell's genome has more bound chromatin than normal, and this plus some chemicals might stabilize the DNA.  Alternatively, some organisms like R. durens have a remarkable ability to maintain their genome (they are tetraploid I think).  Perhaps one could isolate and purify a suite of stabilization proteins from them.

I'm clueless when it comes to chemistry.

Mac

Nathan McCorkle

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Feb 25, 2009, 5:27:41 PM2/25/09
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Can someone post the difference of mini prep, chloroform/phenol, and doing something like a detergent lysis with proteinase K, ammonium sulfate, and alcohol isolation?
--
Nathan McCorkle
Rochester Institute of Technology
College of Science, Biotechnology/Bioinformatics

Cory Tobin

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Feb 25, 2009, 8:48:53 PM2/25/09
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> Can someone post the difference of mini prep, chloroform/phenol, and doing
> something like a detergent lysis with proteinase K, ammonium sulfate, and
> alcohol isolation?


A mini prep is a method of purifying plasmid DNA from the rest of the
cell, including genomic DNA. There are different methods of doing
this. Most commercial mini prep kits use a silica resin to bind the
plasmid. There is a long list of methods for doing this. One is the
detergent lysis that you mentioned. A few others involve alkaline
lysis, boiling lysis, and polyethylene glycol. For a more complete
list, check out the table of contents of chapter 1 in "Molecular
Cloning" by Sambrook. Amazon will let you preview it...
http://www.amazon.com/Molecular-Cloning-Laboratory-Manual-3/dp/0879695773/ref=pd_bbs_sr_1?ie=UTF8&s=books&qid=1235609773&sr=8-1
Also, the terms Midiprep and Maxiprep are basically the same thing as
Miniprep except they refer to doing it in larger volumes. Mini < Midi
< Maxi


Phenol chloroform extraction is a method for separating DNA, RNA and
protein. I wouldn't exactly call it "purification" since more steps
are required to prep your DNA or RNA before doing any downstream
reactions with them. It simply allows one to separate the DNA from
the RNA from the protein. Wikipedia does a decent job of describing
this method...
http://en.wikipedia.org/wiki/Phenol-chloroform_extraction


Ethanol precipitation is a method for concentrating nucleic acids and
polysacharides. Basically the ethanol (or isopropanol) allows the
phosphate backbone of the nucleic acid to react with cations in the
solution, causing it to precipitate. Usually this method is used
after separating the DNA from the RNA and protein (phenol chloroform
extraction) since both DNA and RNA will precipitate in an ethanol
precipitation. For example, say I wanted to PCR off of the genomic
DNA of my epithelial cells. After collecting the cells I would lyse
the cells, then do a phenol chloroform extraction to get the DNA, then
precipitate it with ethanol, dry it out in a vacuum, then resuspend
the DNA in a tris buffer. I would then use that solution as the
template in the PCR reaction.


Ammonium sulfate precipitation is used to separate different proteins.
Since different proteins precipitate in different concentrations of
salt, this can be used as a very crude purification method for a
desired protein.


-Cory

Dan

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Feb 25, 2009, 10:35:33 PM2/25/09
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http://www.biomatrica.com/dnasamplematrix.php?gclid=CPW7vsOf-ZgCFR0Sagod7A8pnw
http://www.genvault.com/html/products/gentegra.html

I know that if you aren't too picky about yield, DNA in TE can be
blotted onto filter paper and shipped with no problem. Phage can also
be mailed that way. (There are some amusing old anecdotes about labs
that were stingy in sharing phage strains and other labs sending them
lots of letters in the hopes that the return letters would be
contaminated with the phage they were looking to get.)

As for RNA, that first link also claims to have an RNA version of
their magic paper.

Proteins - most are a no. Drying tends to terminally denature a lot
of them. (Obviously - see phage - some can survive this but there's
no a priori way of knowing which ones.)

Cells - no, unless you're shipping D. Radiodurans and you'd still have
to deal with contaminating organisms coming from spores on the paper.

Dan

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Feb 25, 2009, 10:49:08 PM2/25/09
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Quick addendum to the last message, I've never used the products in
those links so caveat emptor.

@ Cory - good summary. I'd add the following:

Minipreps - the easiest, fastest way to get high purity DNA. Tends to
be expensive since it's kits. Don't buy Qiagen. There's plenty of
other kits out there just as good that cost 2/3 as much. Also, I
understand you can regenerate miniprep columns by storing them in 1M
HCl to depurinate any residual DNA on them.

Phenol/Choloform/Isoamyl alchohol / Ethanol precip - I lump those two
together since the ethanol precipitation is usually used at the end to
bring the DNA into a smaller volume. The phenol/chloroform/ISA
removes most of the proteins from the DNA. It's cheap in bulk since
the chemicals are fairly inexpensive per prep. However, the chemicals
aren't cheap or easy to get in the first place. Also, phenol is
extremely toxic - probably the most dangerous chemical in most molbio
labs. Chloroform is carcinogenic. Isoamyl alchohol - probably bad
for you as well. All 3 stink and require a running fume hood if
you're working in any significant quantities. Also, the DNA you get
is rather low quality - lots of contaminating protein, etc. Also, it
is easy to oxidatively damage the DNA if the phenol is old. I've done
thousands of phenol/CH2Cl2 preps in the past and if I never have to do
another, it'll be too soon.

Ammonium sulfate can actually be used as DNA prep. I used a genomic
DNA prep kit once that used the ammonium sulphate to knock the DNA out
of solution and leave the DNA behind. It worked surprisingly well.
However, you still have a lot of protein left at the end.
(incidentally, ammonium sulfate is a great way to concentrate proteins
and store them in a stable form. It's an old technique that's kind of
been forgotten. Ask me some other time to go into more detail.

The last way is a cesium chloride prep. You fill a tube with a CsCl
(used since cesium is very dense, the Cs Cl salt in solution feels
like liquid lead) gradient so that the solution goes from less dense
to more dense from the top to the bottom of the tube. Apply your DNA
and spin in an ultracentrifuge at several tens of thousands of Gs
overnight. Everything gets separated by density. Your DNa will come
out in a fantastically pure band you extract by shoving a hypo needle
through the tube side and sucking it out. I've never done it
personally but have seen it done. It's a dead technique since it's
horribly time consuming, requires an ultracentrifuge and is very, very
expensive.

Dan

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Feb 25, 2009, 10:49:14 PM2/25/09
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Nathan McCorkle

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Feb 25, 2009, 11:03:34 PM2/25/09
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Is there a wiki page that we could list archive this information? and add prices? I volunteer to add and tend pages.

John Cumbers

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Feb 27, 2009, 1:39:33 AM2/27/09
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On the subject of sending proteins out... I recently received a free Phusion polymerase sample from NEB: http://www.neb.ca/detail.php?id=F-120&ven=NEB  It arrived in a jiffy bag, no dry ice or insulation, which surprised me.  It says to store at -20 (regular freezer temp) I've yet to try it, but I assume they wouldn't be sending it out like that if it were not up to it.   I think Jim H has noted about storing enzymes in high glycerol in order to keep them at room temp, and also that company in Reading UK was mentioned before that ships dry enzymes.

cheers,

John
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