If you want off the shelf pcr-able genomic DNA from meat, why not follow the Dolan DNA Learning Center's Human Cheek cell DNA extraction protocol written in the mitochondrial DNA kit they sell through Carolina Biology Supply Company. Replace proteinase K with McCormick meat tenderizer enzymes and then do an isopropyl precipitation. Using a prescription drug as a reagent is even less accessible than purchasing the reagent in the first place.
Saline, a centrifuge (hand crank? Dremel?), tenderizer, etc all fairly accessible. If you do want to make it more off the shelf why not process plants instead. Just soap, salt, and alcohol is all you need for a PCR-able crude genomic DNA extract. Meat is a little bit trickier but the tenderizer enzymes (papin proteases) will handle the tough connective tissue bits quite well. I did a science project when I was a wee lad and extracted genomic DNA from chicken liver using household products. It worked but never did any pcr on it since this was in the 1990's and PCR machines were not easily found.
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
Saline, a centrifuge (hand crank? Dremel?), tenderizer, etc all fairly accessible. If you do want to make it more off the shelf why not process plants instead. Just soap, salt, and alcohol is all you need for a PCR-able crude genomic DNA extract. Meat is a little bit trickier but the tenderizer enzymes (papin proteases) will handle the tough connective tissue bits quite well. I did a science project when I was a wee lad and extracted genomic DNA from chicken liver using household products. It worked but never did any pcr on it since this was in the 1990's and PCR machines were not easily found.
Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC
Plant Biotech R&D
From: leaking pen
Sent: 1/16/2015 8:21 AM
To: diybio@googlegroups.com
Subject: Re: [DIYbio] DNA extraction and DNA Barconding
the specific restriction enzymes used cut known spots that occur in pretty much all coding, and the distance between those codes, and the percentage of fragments that fit it, are actually very revealing to identify species.
On Thu, Jan 15, 2015 at 9:34 AM, Otto Heringer <ottowheringer@gmail.com> wrote:
Hello people,--I'm planning to perform a DNA extraction to do DNA barcoding analysis with a sample of food (bovine meat, I suppose). The DNA extraction part is for a public demonstration of DIYbio and a "first experiment" introduction - one past thread on this list helped figure out this.The objetive is do a DIY extraction using common commercial products and a "normal" extraction using standard lab reagents, just for comparison before the public demonstration.Cathal's blog have the best source of information regarding this subject I found so far - specially where to find some "ingredents".I'll look for genomic DNA instead of only plasmids (I'm using this protocol), but the problem of where to find some reagents persists. The only one that is hard to obtain is indeed Tris.My plan is try to use drugs like Toradol that is usually sold with "Thrometamine", what appears to be another name for our well known Tris. I don't know if the information about the % of thrometamine on the drug will be avaliable, But I'll try do something.
Do you know other better options? Maybe a more avaliable compund that might substitute Tris?And about barcoding, I see that there is a still open debate about the use of barcodings because of the supposition that it might "replace" taxonomy. Some say it is only useful for "biodetection" of some species and that taxonomy need more complex work.But my point is: if barcoding is only useful for biodetection, so why is needed to use only a particular DNA region present in all species? If the biodetection was the only goal, wouldn't be better compare whole genomes and trace a map of DNA disparities between all species? Of course, taking in acount the crescent feasibility ($ and time) of whole genome sequencing.This way just a PCR and gel would be needed - and not sequencing for every barcode sample.I know some of you have some experience on the subject. What do you think about it?Otto
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