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e-DNA Sampling For Sasquatch


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Catmandoo

Hiflier, you have been busy. I have not put much thought into the e-DNA effort because I assumed the cost to be out of reach and historically, the DNA faking and hoaxing in the past has created a huge hurdle to climb over.

 

Those $15 test kits from Smith-Root are a tease. Smith-Root has lab services available for e-DNA by partnering with Precision Biomonitoring. Cost is unknown. Smith-Root is focused on aquatic species. No mention of land animals. I am working on this:  I would submit a sample in the way of searching for a large member of the Squatchmonidae family which is a member in the order Squatchmoniformes.  That would keep them guessing......for a few seconds. Lab services are a big unknown. 

 

Peristaltic pumps contact the sample. Vacuum pumps are less money and don't contaminate the sample. I have 2 versions of Mityvac. American made and other. The gauge is not calibrated and does not have to be. We don't have vacuum pumps. Vacuum exists in space. We have 'negative pressure' pumps.. I used to vacuum bag oak veneers using resorcinol glue doped with about 10% alcohol. I used the Mityvac hand pumps. Strong hands are needed. I could pull down to 23 inches mercury negative pressure and hold that overnight. I put several loops in the suction tube and flashed off some alcohol which settled in the bottom of the loops as a liquid. The pump does not have to be able to hold negative pressure. Just pinch off the tubing.

 

Liquid could be sucked out of a track with a hand held pump and the $15 test kit.  I would suggest food grade silicone tubing ( after the test kit ) since you can do a thorough cleaning / sanitizing on it. 

Dirt is a few more steps and equipment. For dirt, I would use a hygenic scoop. Place sample in bowl. Add distilled water  Proportions are theoretical. Perhaps 50-50.  Blend well.  Place bowl in vacuum bell.  Pull negative pressure and flash off the water through test / sample kit. The Smith-Root test kit is rated to 12 psi and a negative pressure of 24.4 inches of mercury.

 

The factor of human contamination has destroyed efforts in the past. What if you could uniquely 'mark' your DNA.   I believe that I can 'mark' my DNA. Theoretically, lets say I contaminate an empty collection object like a bowl, cup or plate.. I whip out my UV-C light(s) and bombard the collection object. Dose and time are factors. Thymine in my DNA is damaged. Uracil in my RNA is damaged. Would the damage be an identifier for me in a sample with multiple DNA traces? Maybe. UV-A and UV-B are worthless for sanitizing so don't buy that junk on ebay and Amazon.

 

I have all of the toys and dollies including the expensive googles to protect my eyes from UV-C.  UV-C is not on Earth. It is in space and gets filtered out by the atmosphere. UV-C is dangerous and will blind animals and humans.

I have too many irons in the fire currently. I will have to suck on some mud in the near future.

 

Edited by Catmandoo
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hiflier
Posted (edited)
1 hour ago, Twist said:

.....Serious research needs to be done on the subjects of A) Can EDNA produce results that could show the right gene.   B.) Who and how can the gene be identified by. 
 

Both questions that should be answered before a grassroots movement has ppl spending money on collection kits.  

 

You are spot on, Twist! So trust me when I say that THAT is exactly why I've been emailing scientists...because I have needed those very questions answered !! Especially with regard your "A" question. Because I knew that unless I got at least that answer then there would be a big hold on what I've been trying to do here. The short of it is I got my answer and am now waiting for a second opinion from another source. First source? Told me my concept would be a "great study" but mentioned blood, bone or tissue as samples as being the best to test from (we already know that).

 

Of course I was presenting the hypothesis of looking for those pesky NOTCH2NL genes I've been yammering about for the past four months. But my source did on to say that a well-designed DNA project, the collection of water samples could reveal an unknown primate. Also that there is a thousand times better chance with mtDNA than nuDNA to capture novel primate DNA because there's so much more of it. My NOTCH2NL genes apparently are nuDNA genes (chromosomal) which I need to do research more on (moron...see what I did there? ). In retrospect, maybe "more of" may have been more personally prudent? ;) 

 

@Catmandoo Thank you for bringing in the good stuff! Can you take over the helm here while I go look for labs? Especially for where a reasonably priced portable vacuum system could be had  :) 

Edited by hiflier
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Catmandoo

I would not take electrically powered pumps into the field unless I used an ex-friend to carry the batteries.  I have strong hands so I can pump like a maniac to pull good negative pressure with my Mityvac.

 

The DIY vacuum set up will be sourced from various vendors.

The choke point is still the test kits / lab services.

 

Mityvac       about $40 at big box stores. Various 'kits' are available but one just needs the basic kit.

food grade silicone tubing      buy as surplus on ebay

vacuum bell jar    cheapest will be on ebay and Amazon. Small is better and a base board with sealing gasket is needed.

 

BTW, sending in hair and turds does not work.

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hiflier
46 minutes ago, Catmandoo said:

BTW, sending in hair and turds does not work

 

Evidently, nor does DNA ;) (just kidding)

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BlackRockBigfoot
57 minutes ago, Catmandoo said:

BTW, sending in hair and turds does not work.

Story of my life...

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hiflier
Posted (edited)

@Catmandoo How does one incorporate a bell jar in a water system? Air? no problem. But Water? Does the water enter the jar from the filter and exit to the pump with always at least some air in the jar? Is it so a more consistent negative pressure can be applied to the system without the drastic and unwanted stops and starts that just a plain hand pump would generate?

 

I could see negative pressure in jar pulling in water that finds its way to the exit port of the jar. The water in the jar would trap a pocket of air which would cushion the effect of hand pumping. As long as one keeps pumping so that the negative pressure in jar keeps pulling water smoothly through the filter than ya got it licked :) Just keep an eye on the water level in the jar and , accordingly, pump more or less to maintain the water level/flow from filter.

Edited by hiflier
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Catmandoo

I would not use a vacuum bell jar for a sample from standing water. The pump would be used to suck water through the sample filter. Those kits from Smith-Root have a suction hose. The filter assembly can handle as much negative pressure that you can generate, about 24 inHg.

 

I would try the vacuum bell jar for a 'dirt' test.  A slurry may have to be more than a 50-50 mix of dirt and distilled water.  Perhaps 90:10,  90% water to 10% dirt. Soils vary. Western Washington soils have varying amounts of clay which is always a problem during mineral prospecting. Many trials on slurries would have to be made to determine if flashing off water vapor  under negative pressure scenarios would produce enough liquid to be viable before field use.

 

Hiflier, find a lab that will process the Smith-Root kits regardless if the target animal is aquatic or terrestrial.

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NatFoot

Recommendations on gear set ups should be coming from professionals...hopefully folks from the identified lab that will be accepting our samples.

 

No disrespect, @Catmandoo.

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hiflier

Most of the equipment recommendations come from the USGS .pdf that I posted. But specs are specs, and as long as the proper hg vacuum pressure can be somewhat consistently maintained and the tubing doesn't collapse between the filter packet and the pump then I think equipment sunstitutions would be okay to make. But I agree, NatFoot, some input from a lab would be beneficial.

 

Cat, one would think that any lab could open the Smith-Root filter housing and extract the filter material. I like the idea of using that product because the alternative would be the one from Biomeme, which doesn't look as field friendly, or the traditional method of folding and placing a loose filter membrane into a tube pre-filled with ethanol.

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wiiawiwb
MODERATOR

Here is an e-DNA aquatic kit for collecting a sample from JonahVentures. It uses a large syringe to push the water through the filter.

 

https://jonahwater.myshopify.com/

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hiflier

Thanks, w, that looks great for fish and algae and maybe down the road the company will use any profits to expand what they can test for (mammals?). I assume when one buys the kit that the cost for analyzing the sample is included? They didn't say one way or another so, if true, it's a pretty cheap way to assess someone's pond or stream. It would be an interesting gift for a young person to get them more interested in the environment.

 

The Diet kit is interesting in that it supposed to detect what animals eat by testing fecal material. It looked like the only candidate was the Prairie Chicken but I would figure fecal material from anything could be sampled. Speaking of which, 50-60cc's of test water is a pretty small volume. Makes me wonder if there SO much DNA in a pond that that's all that's needed.

 

   

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Catmandoo

NatFoot, no problems.

The suggestions from Smith-Root are their field testing combo packages that are over $5,000 and top of the line backpack unit @ $6,000.

The $89 unit from Jonahwater appears to have some sample processing included with the purchase price. The $15 kit from Smith-Root is an item to check out. Lab fees are not posted on their site.

 

I did not waste any time on the Disotel song and dance routine. Perhaps someone can review his comments to determine if he stated what equipment / lab should be used. Might have a Hollywood certification.

Citizen science is difficult. The DNA arena has been trashed by the hoaxers. Finding a lab is an uphill battle.

Edited by Catmandoo
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hiflier
34 minutes ago, Catmandoo said:

Finding a lab is an uphill battle.

 

So I'm finding out, though I do have a couple of more emails out. Like Norseman, I,  and gigantor said, this is going to take some time. In a way that's okay because it gives us time to get our logistics questions ironed out

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NorthWind
14 hours ago, hiflier said:

 

 

@NorthWind and @BlackRockBigfoot Is this maybe what you are looking for? Soil Sampling.pdf 

 

Then you, me, and everyone need to know the general procedure that has to happen once samples are delivered to a lab. I also think this video shows why the soil samples  from the OP nesting site were so expensive to test:

 

 

 

Hiflier, the .PDF, while giving good tips on the collection, doesn't really fully apply, as they are looking for specific fauna in the soil, suck as mites and springtails. We're looking for something just a tad bigger :-) Anyway, I will check out the video when I get some free time, something which I don't have a lot of these days it seems.

 

And then, not to throw a monkey wrench into things, (pun intended) but what if BF is not primate at all? We are making (an educated) assumption. Seems like it should be. But who knows? 

 

Still, this idea seems kind of exciting.

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Madison5716

I'm in. This fall when we find prints (yes, I am that confident), NorthWind and I will contribute. Sounds like an awesome plan!

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