A commenter over at my blog has asked about leads to information regarding sasquatch genetics. Who did the work, their results, GenBank numbers and the like. Your assistance would be most appreciated.

There is none. All the data remains just anecdotal, nothing is published to my knowledge, and nothing has turned up pointing to anything but human contamination (or something which is virtually human??? ).

Apeman

Since moving to the northwest United States I have had an on again, off again, interest in sasquatch from the occasional stories I hear. I have been working in the field of genetic research for some time, which leads me to the following question: Why hasn't hair been been used more often to look for genetic evidence of sasquatch's existance?

If this creature exists, it's mitochondrial DNA should be very distinct. Using modern DNA lab techniques, extraction, PCR amplification and sequencing of hair samples is a rather straight-forward process (this is, of course, assuming the geneticist knows what they are doing). Contrary to popular belief it is not necesary to have root tips. Hair shafts are sufficient. As a rough estimate, the cost of verifying sasquatch DNA from a hair sample should be about $500. At first glance that may seem like a costly sum, but proof would be indisputable, and therefore priceless.

Has little DNA analysis occured because scientists (who are mostly academics) are reluctant to risk their reputations on such research? Is it the cost? Or is there simply no place a saquatch researcher can send DNA for analysis? I'd appreciate your thoughts.

Bluegenes;

From what I've read, we have collected samples of sasquatch hair, and could get very limited DNA examples from it. Even if we obtained a full strand of Sasquatch DNA and analyzed the hell out of it, what would we compare it to? We'd need an actual sasquatch specimen to verify that the DNA is from a sasquatch, and at that point we'd know sasquatches are real and wouldn't need the DNA sample. It's a catch 22 scenario.

I think hair hasn't worked out because of the lack of a medulla in most credible hair samples, i believe, although i'm no expert on this.

I was talking to someone yesterday who is doing their masters thesis in molecular phylogenetics, and he commented that he thought it would cost in the ballpark of $5000 to have a very fresh (or quickly freeze dried) hair with a follicle identified as primate or non-primate, and if you were lucky, to hone in on the type of primate. For about $15,000, you could get multiple sequences produced, as well as a phylogenetic tree.

That was just an 'off the record and off the cuff' guesstimate, but it's an order of magnitude more than your number..

I think the problem right now is that there isn't any good material to work with. No tissue samples, and the hair samples so far have been fairly useless for extracting DNA.

I think this is the wrong premise. A lot more has occurred than most are aware of, but it generally hasn't yielded good results. Reputation preservation seems to be a non-factor in my experience and costs are also rarely a consideration if the samples get into the "right" hands.

Robo- That guy must be talking Jamaican dollars?

Apeman

Heh.. nope, but i have no way of judging it.. it was just a whole bunch more than BlueGenes' number. It sounds exorbitant to you?

I think it may have been in the context of commissioning a study at a university, rather than the actual cost of materials and a lab tech's time, but i'm not sure. I'll ask.

First of all, I'd like to apoligize for perpetuating a topic that has been previously discussed. I am new to this board. It was only after I wrote my first reply that I searched the boards for similar topics. It was then I discovered much discussion has been devoted to DNA analysis. Nevertheless, I do think DNA can and should be used as a tool to compliment other previous and ongoing research.

In regards to concerns that one would need an actual sasquatch specimen to compare a DNA sequence to, and hence, creating a catch 22. I can certainly understand this line of thought. During my graduate school course work I took several classes on phylogenetic analysis and heard students, new to the field, voice this concern. But respectfully, phylogenrtic analysis doesn’t work that way.

Let me give you an example: Lets suppose we know the sequence of a gene found in both humans and dogs. When we compare the human sequence to the dog sequence we find that 60% of the human sequence is the same the dogs. Next, lets assume that up to this point in time chimps have not been discovered. We have a few photos, some sightings, hair samples, etc., but no whole specimens (live or dead). We can guess from the photos that chimps are more closely related to humans than dogs, but is that really the case?

Using the chimp hair samples we can sequence the same gene we have previously sequenced in humans and dogs. We then compare the three against one another. As it turns out the chimp sequence has a 90% similarity to the human sequence but only a 70% similarity to that of the dog. From this, the conclusion is simple. Chimps are more closley related to humans than dogs.

To expand on this idea: there exist public databases that contain literally millions of sequences from many thousands of organisms. Using these databases we can compare the chimp sequence to all other described species (in which that particular gene has been sequenced) and pin point the chimps evolutionary location in the animal kingdom. So, you don’t really need an actaul sasquatch specimen for comparative analysis. I hope this clears this aspect of DNA analysis up?

BlueGenes

Concerning lack of medulla as a reason that DNA can not be extracted from sasquatch hair. Lack of medulla has long been suspected as a reason that DNA could not be extracted from some hair samples. However, an extensive study, published just this year, has shed light to the contrary.

It turns out that absence or presence of medulla is not a factor in successful DNA extraction and subsequent amplification. Other factors were found to have large effects, however. Most had to do with the quality of the starting sample. For example, hair that has been exposed prolonged and repeated wetting and drying (rainfall) or UV light (as in sunlight) did not work well. The age of the hair was another factor. Treatments to the hair, from dyes or preservatives also resulted in poor DNA amplification.

So what does this mean? If one finds a saquatch hair sample that has been exposed to the elements for a long period of time, then collected and put into a moist plastic bag, then stored in a musty desk drawer and handled repeatedly by friends and acquaintances for many months or years, odds are the hair sample will not yield useful DNA. (Please understand, I am not trying to paint sasquatch researchers as bumbling amateur scientists who can’t collect a proper hair sample. I honestly do not think that is the case. I’m simply trying to paint a worst case scenario. )

If a hair sample is collected with a gloved hand, air dried if necessary, stored in a paper envelope or container and touched and handled by few or no people, useful DNA can be extracted.

DNA protocols have come a long way in just the past decade. What was thought to be a sample that was too small or too degraded just five years ago, is sufficient today. And the future looks even brighter.

BlueGenes

About the cost of a DNA analysis: Just this week (using grant money I will admit) I bought a supply of kits and chemicals necessary to extract, amplify and clean DNA for sequencing. I have done some additional math and my $500 figure seems about right.

As I said in a previous post, DNA analysis has come a long way in a short period of time. Costs have been reduced considerably in the past few years. It’s true that five or ten years ago such an analysis might cost several thousand dollars, but that is no longer the case.

With the explosion of genomics in the past 20 years even labs in some small colleges have the necessary equipment to do DNA sequencing. Any major state or private university will certainly have several labs with the capability. I have even set up a small lab in my home to do DNA work (it allows me to work from home one or two days out of the week).

A caveat: If a good sasquatch DNA sequence was found, it would need to be verified by another independent lab. Additionally, a very detailed phylogenetic analysis would need to be constructed and a peer reviewed, journal quality paper written and published. All of that is time consuming, and hence expensive. However, if a good sequence were found I have no doubt some geneticists would jump on the chance and do the work pro bono, simply for the notoriety.

(Enough writing for now....I need to get back to work and earn my salary!)

BlueGenes

Thank you BlueGenes Very informative. It helps some of us understand the subject better.

BlueGenes: I'm only slightly versed in the kind of genetic work you're talking about: I kinda know what PCR is, but that's about it.

Simply out of curiosity (I'm a geek that way), what kinds of equipment and supplies does one need to do sequencing? What kind of work is your home lab capable of? How do tools like BLAST fit in? What can sasquatch enthusiasts do to improve their collection techniques? (I bet more samples collected fall into a scenario like your example than we'd care to admit.)

Setting aside the scarcity of samples, either viable or not, what can enthusiasts do to help someone like you have a go at this?

A great topic for sure. And thank you for the updates.

I studied Genetics a bit in college, but than that was in the dark age of genetic research.

Much has changed........... in fact most has changed.....

Thigmo: To answer your questions:

Obtaining a DNA sequence is a four part process.

1. The DNA is extracted from a tissue (Hair, skin, leaf etc.) sample. This requires about 5 different chemicals and a centrifuge.

2. A PCR (polymerase chain reaction) is performed. Currently, in order to obtain a DNA sequence read, one needs multiple copies (many 1000's) of your gene of interest. Raw DNA that has just been extracted from a sample usually only has one, a few or in some cases a few hundred copies of any one gene. A PCR creates literally millions of copies of the gene you want to sequence. This is done by adding about 5 or 6 more chemicals as well as "primers" (crudely described, they are enzymes specific for the gene you want alot of copies of). This chemical mix is put into a machine, called a thermal cycler, which heats and cools the reaction mix many times to specific temperatures at specific rates (temps and rates that make the primers work).

3. Now that you have a test tube with millions of copies of you gene of interest it needs to be cleaned. The cleaning process removes all of the previous chemicals you have added and leaves you clean copied DNA in a buffer.

4. The copied DNA is sequenced in a sequencing machine. The result is a read (sequence) of your gene of interest. The sequence will be composed of a string of nucleotides (A's,C's, G's and T's). The sequence length will vary depending on the length of your gene. It can be short (<100bp) or quite long (currently sequencing machines can only provide reads about 1000 nucleotides long).

I could also add another step to this process. Recently repair enzymes have availible at a reasonable cost. These enzymes can repair DNA that has been degraded because of age, UV damage etc. They won't do miracles, but they are good at mitigating minor DNA damage.


In my lab at home I can do steps 1-3, providing I'm doing a few samples. If I'm doing a large number of samples, or need to "tweek" any part of steps 1-3 I'll work in my lab at the University. The cost of the equipment for varies. A crude lab can be set up for about $1000-$3000. The lab equipment at the university, as one might suspect is worth much more. I do have access to a sequencing machine, but honestly it's real pain to set up and run. I usually send my cleaned DNA to a high throughput sequencing machine housed in another building on campus. There are also several private labs around the country that will sequence cleaned DNA. SEquencing machines are very expensive. An old used machine might be had for $100,000 while a new high speed machines run in the millions.

I honestly wouldn't recomend that someone try to set up a private DNA lab in their home unless they have had some experiance working an active lab. The techniques are certainly learnable, but it does take time and a great deal of practice.

I hope this helps satisfy your "geek" curiosity!

As far as hair collection techniques, this is the protocol I would use:

1. Collect the hair sample using sterile latex gloves. These gloves are cheap and can be found in most drug stores.

2. Place the hair (wet or dry) in a clean, previously unused paper container (an envelope or cardboard box).

3. If the hair is wet, allow it air dry. Do not use a blow dryer or an oven!

4. As stated previously, store the hair in clean paper container. I would not advise freezing the hair unless it will not be analyzed for a long period of time (over one year). Ideally samples stored for a long period of time should be frozen in a -80 C freezer, but an ordinary household -20 C freezer will do. The reasons I don't recomend freezing are thus: Unlike skin or blood, hair (and the DNA within it) will not degrade at a fast rate. Seceondly, during shipping to a lab the sample is likely to thaw. This will cause condensation and perhaps lead to contamination and degradation of the DNA. Addtionally, the temperature fluctuations are not helpful to DNA preservation.

5. Store the paper container in a dark area that will not experiance wide fluctuations in temperature or humidity. An air conditioned and heated room is good. A moist basement is not.

6. Send the sample to a lab that will do direct DNA sequencing. Other methods, such as DNA finger printing and blots are fine for other applications, but for species determination and phylogenetic analysis, direct sequencing is currently the most robust and unambiguous method.

The lab should also agree to BLAST any sequences they obtain. Blasting works this way: The US gov't owns and maintains a "bank" of gene sequences known as GenBank. Other banks exist around the world, but GenBank is the depository most scientists use. Actually, most peer reviewed publications require the scientist to deposit their sequences in GenBank before publication. As a result GenBank contains millions of DNA sequences. Using the GenBank webpage anyone can "BLAST" or compare their sequence to all other sequences in the database. One of the things the BLAST program does is return a spreadsheet detailing what other sequences match yours, using a percent score. All sequences submitted to GenBank must contain, as part of the file, a name of the species sampled. At this point further comparitive analysis can be done.

Hope this helps.

BlueGenes

I have read that the mitochondria and the genes are often degraded in the process that the body uses to make hair. I have also heard that the mitochondrial DNA is primarily found only in the medula. After reading of them finding hair that was thousands of years old and getting useful data it made me suspect they weren't actually extracting the DNA from the hair efficiently. Do they use enzymes like kerotinase that break down the kerotin in the hair before they attempt to extract mtDNA from it. Hair seems rather dense to allow large molecules of DNA to diffuse through it.

Check with Todd Disitell (spelling). He's done a couple of DNA tests.

Great post - thanks...

BTW I emailed the guy giving the $5000 quote but haven't heard back

Proteinase is the enzyme used to break down the hair shafts for DNA removal. As I mentioned in a previous post, the presence or absence does not seem to be a factor in whether DNA can or can not be extracted....it's other factors such as the original condition of the hair sample.

BTW....my figure was $500 to obtain a sequence...not $5000. If someone were to charge $5000 they would be making a very healthy profit for their work!

BlueGenes

I'll tell you guys what........I'm willing to try getting a DNA sequence from a sasquatch hair sample, once, bro bono. I would like to hear the circumstances of the hair collection first, however. My work should be slowing down for a short time next month and I could fit it in at that point. Please let me know.

BlueGenes

Henner Fahrenbach had curated some samples for purposes of microscopy at one point. I don't know if he still has them, or if they have been stored in such a way that wouldn't contaminate or render useless any further testing.

He doesn't post on this forum, and I haven't contacted him in a few years. I'll reach out to some folks who know a good deal more about it than I do.

I sent him a hair sample ( Fahrenbach) awhile back and he was prompt and gave me the most detail. Turned out to be probably skunk. I 've hear that he might have as many as 5 hair samples from good encounters. It's worth a try. I have his e mail and address if you want it.

Bluegenes,

Thank you for your offer and your technical advice!!

Hopefully, someone will be able to take advantage of your kind offer.

For what it's worth, i got a response from the guy. I think the first quote was somewhat tongue in cheek (hard to tell over email), as the cost for him to switch his master's thesis over to the analysis of a Squatch hair sample :rolleyes:

Anyway, this is what he said:

"$500 might be in the ballpark for one or two sequences. If you want to do a proper taxanomic inquiry you should sequence multiple sites and then pay someone to search through the literature and genebanks and generate a phylogenetic tree. And again, if you come up with "unknown species" it doesn't mean its a yeti, it could be a known species that hasn't been sequenced, so one would need to go sequence those...

So $500 maybe for a sequence, yes? but lots more for a proper investigation.

It would cost under $100 for the lab materials (if purchased in bulk, which most labs would), but the labor costs are what bumps it up. It could take me 1-2 days to sequence DNA (mostly waiting for various reactions honestly)."




Anyway, BlueGene's offer sounds great... if only there were more good hair samples.

Thanks for the response Robo. It's much appreciated. I'm glad your friend agrees that the $500 figure isn't out of the ball park. I was figuring the genbank search and phylogenetic analysis would be done mostly pro bono. Some evolutionary scientists I know might do such work just for the challenge and novelty. I certainly would. I still disagree that you need a confirmed BF to compare a sequence to. My line of reasoning was hammered into my head by more than one professor in school. (Maybe I'm now the product of that cruelty ) Tell your friend I said thanks and good luck with his work....I know from experiance how hard it can be.

I don't have any plans to contact Fahrenbach for his samples. Since they were not his originally, I doubt he would give them to me. Additionally, I would be receiving them third hand....... down the line someone could use that to poke a hole an any analysis. I won't rule out using samples from him, but right now I'm not all that keen on it. Hopefully someone with a good sample will see this thread and we can go from there. We'll see...............

BlueGenes

As I said a week ago here...and probably a dozen times in earlier threads.





Apeman

Since the technology seems to be just waiting for a decent sample to work on, the interesting question, then, is WHY there is little or no good hair evidence. After footprints, it would seem like the most likely physical evidence to be collected.

It's interesting that the prevailing attitude that i've noticed is that hair evidence isn't of much use. Most recently, this view was given in the last BipCast, IIRC..

Perhaps that view is outdated, and it's time to really make a concerted effort to collect hair samples?

That's a good question Robo. I wish I had a good answer. I suppose it could be one of many reasons. Perhaps hair is not sent to labs because of expense. Pehaps it's sent to the wrong people (those who look at it's morphology only). Maybe most people think DNA analysis is a bunch of nonsense. I just don't know.

Still no responce to my offer analyize a sample in my lab. Too bad. I think I'm starting a get a definate opinion about

I suspect that one reason we have so few good hairs for examination is the provenance of the sample. Unless you see the hair pulled out of The Big Guy as he goes over a barbwire fence, it is hard for a lot of us to pop with the money for a DNA test. If the hair is found on it's own, your not sure where it comes from for sure. Even if it is quite unusual, $500 is a lot for an unknown sample that could very likely come back as a regular farm or woods animal.

For example, a year or so ago, there was a report of a bigfoot leaning over a fence looking at a kids playground in upper Canada. A hair sample was found along the fence. But turned out to be cattle or otherwise known animal. Now it could have been a legit sighting. But in fact did not leave hair to be examined.

So it is difficult to find a good sample and know exactly where it came from. Even if you see a 'foot go running off into the woods. Then as you are trying to find it, you come accross some hairs in a branch - you can still not be 100% sure of its origins. More likely you will send it in for the cheaper microscopic exam than the cost of a full DNA test.

It's a bit difficult to imagine a situation in which you could be certain it was BF hair?

Perhaps if you ran up and pulled a handful out of the big guys chest.............

Hey, that could be a jerky commercial..........

It's correct that a microscopic exam is far less expensive, faster and easier to do than a DNA analysis. Perhaps things will change in the not too distant future. My onlu comment is: A microscopic exam confirming or disproving a sasquatch hair sample will never be excepted as "proof" by the thinking public or science. Microscopic exams are not in the same league as reading chicken entrails, but that's how most scientists will see it (be that good or bad).

BlueGenes

I agree BlueGene that microscopic examination would not be acceptable as proof. But could stand as the first step twords a DNA test. If a micro exam comes back as unknonw primate, then by all means it goes to a lab and you start planning on 2 months of eating Top Raman noodles to pay for it. If it comes back as bovine, then it's only 3 weeks of Top Raman for the microscopic look. The other advantage of a microscopic exam is that it does not destroy the sample and allows further testing of the original sample.

You make a very valid and smart point Bill. Thank you for adding it!

I've read through these posts but do not see a place to send our samples for analysis.

Please advise if you know of a place that will do the dna testing that is necessary to prove that we do indeed have what we think we do.

Thank you for your help!

Do you have any photographs of the hairs?

SY.