What genetic modifications can one make at home?

[Andrey_]

Can one make a genetically modificated plant or animal "at home"?

[Leader Bee]

I don't know if it counts but over time I suppose you could use selective breeding (is that the right term for plants?) to produce a strain with properties you desire - I.E larger/More fruit or something that simply grows faster.

 

You would of course need examples of these plants to breed into something where these particular properties were more prominent.

Edited January 11, 2012 by Leader Bee

[Tres Juicy]

I don't know if it counts but over time I suppose you could use selective breeding (is that the right term for plants?) to produce a strain with properties you desire - I.E larger/More fruit or something that simply grows faster.

 

You would of course need examples of these plants to breed into something where these particular properties were more prominent.

 

 

Cross pollination?

 

People regularly do this with cannabis at home (or wherever).

Edited January 11, 2012 by Tres Juicy

[Leader Bee]

No I don't think it is necessarily cross pollination, because that would involve using seperate species to germinate a new plant with the desirable properties. Using pollen from the same species but from plants within that stock that have been selected specifically because of their desireable traits is more what I meant.

[Tres Juicy]

No I don't think it is necessarily cross pollination, because that would involve using seperate species to germinate a new plant with the desirable properties. Using pollen from the same species but from plants within that stock that have been selected specifically because of their desireable traits is more what I meant.

 

 

selective breeding it is then!

[Andrey_]

Actually i was talking about mutants. I came upon some articles like this, where amateurs are making genetic modificated bacterias. And I'm very curious is there any way of making plants or animal mutants in your garage?

[Arete]

Actually i was talking about mutants. I came upon some articles like this, where amateurs are making genetic modificated bacterias. And I'm very curious is there any way of making plants or animal mutants in your garage?

 

In a controlled manner, no.

 

The cited article is nice, but pretty naive.

1) Modifying bacteria is a rather routine laboratory procedure. You can buy a bacterial cloning kit and insert a gene of your choosing using relatively basic benchtop techniques. Doing it at home shows significant determination, but not a whole lot of novel innovation. http://www.invitroge...Brand/topo.html

2) The author seems fairly poorly informed - luminescent tattoo ink already exists for retail sale http://www.crazycham...yartsupply.com/ http://poundedink.com/tattoos-that-actually-glow-in-the-dark/ and genetic modification is unlikely to yield a viable ink. Unlike luminescent pet fish whose whole bodies glow due to the expression of modified luminescence genes, to just make a tattoo glow using GM, you'd need the ink to be alive, or to mutate the host's cells. If it was that easy to modify a human skin cell, many medical conditions would have been trivial to cure for a long period of time.

3) A -80 is a fairly basic laboratory storage device. Even setting up Sanger sequencing will set you back a decent six figure sum (let alone next gen techniques). While it's awesome that civilian scientists are interested in biotech, it's an equipment intensive field and without industry standard equipment it's hard to innovate things industry/academia hasn't already tried.

4) GMO is seriously ethically bound. DuPont spent 2-3 billion dollars generating BT insect resistant sunflowers. When experiments showed that, at least in north America, the BT genes would cross into wild sunflower populations and negatively affect native herbivores, the project had to be dumped. Given the potential impacts of GMO's, they're not the type of thing you want unregulated parties whipping up in their garage - even if they could.

Edited January 12, 2012 by Arete

[Tres Juicy]

Actually i was talking about mutants. I came upon some articles like this, where amateurs are making genetic modificated bacterias. And I'm very curious is there any way of making plants or animal mutants in your garage?

 

Yeah, definitely don't please

 

I don't want enthusiastic amatuers whipping up viable strains of some horrible bacteria in their garage thanks

[Andrey_]

I've heard, but didn't manage to find any information, about one guy, who was making luminiscenting frogs, by ingecting their cavair with smth. Is this method possible or it is fake inf? Is it possible to modificate animals which come from eggs or cavair?

Edited January 12, 2012 by Andrey_

[questionposter]

I forgot what the instrument is called, some kind of incubator, but you can buy it for a ton of money and it will insert custom plasmids into bacterium. There might be a similar things for viruses that you could introduce into your body, but that would never be available to the public.

[CharonY]

Actually i was talking about mutants. I came upon some articles like this, where amateurs are making genetic modificated bacterias. And I'm very curious is there any way of making plants or animal mutants in your garage?

 

The article (and several of similar ones that were published that time) was horrible. And afaik nothing ever came out with that. One of the things that were proposed (melamine detection) were fundamentally flawed. Less due to technical issues but rather due to lack of biological understanding.

 

Technically, low-cost solutions exist to do very simple experiments. However, considering the amount of troubleshooting required and also the constant risk of contamination and other issues make it highly unlikely that an enthusiast will manage to perform successful experiments (I am talking undergrad lab-course level) at home. With determination and signficant time and money investment it is theoretically feasible, however.

 

The biggest risk I see is relatively simple. They may cultivate contaminations that may include pathogenic strains living on skin, for instance to dangerously high titers.

Edited January 16, 2012 by CharonY

[Pickerel]

Sorry to dredge up a dead thread.

I'm going to avoid the article given the reviews, and give my own personal experience:

Modifications are time consuming and tedious to get correct. There's many little fiddilybits one may or may not expect. You will very likely, excluding beginners luck, spend months trying to get one thing or another to work. Even the simplest, regular PCR amplification of a single small bit, may take several tries. Overall, the main first thing you will need to do is read up in primary articles AND books and such on anything pertaining to the group of organisms you want to work with, as well as many other types in case you need to work with multiple to get where you want to go and to get an idea of what it takes. Luckily I prefer yeast, or I'de have a heck of a time ^.^

That said, the field is nowhere near as expensive to get into as the above says. There's a number of things you need, but most can actually be gotten pretty easily. I haven't found a way around the expense of Big Dye, but I try to sequence almost none of what I do. Basically, the trick is to learn a little electronics to keep things working, and to Buy Used. The field has been booming for some 20 years, and is speeding drastically ahead all the time, so equipment becomes obsolete quickly and can often be found cheap. Also, many things can be made on ones own.

You will need:

Thermalcycler. If you are willing to get one online, they can be as low as a hundred bucks for many of the older models. I got my first one on ebay for two hundred plus shipping, and it still works fine.

Organisms: This is up to you what to choose.

Taq Polymerase: The staple of PCR. If you plan to do anything sellable - i.e. if you will make a profit - you will need to purchase special Taq polymerase because it's still under patent (though I've heard this is soon to end! YAYS!). That said, however, researchers are free to make their own using E.coli transformed with the plasmid. Contact your nearest university: most have a biology department, and most of those have someone with the strain. There's a protocol for extracting from said bacteria that might take a day, and took me four tries to get right, but once you have the method down it works well and easily to make vast quantities. If you do this, it will cost close to free - so long as you yourself will not profit from the results.

Centrifuge: You will need a benchtop centrifuge. For most DNA extraction and Taq Polymerase extraction, a typical eppendorf benchtop centrifuge is good. I got mine used online for ~50bucks but the shipping from canada was close to that again. It works well, though, but I had to replace the motor from a scrap one I got for another fifety bucks plus shipping online (which itself had board problems).

Primers: you may have to learn to design primers. Luckily, there's a number of easy to use primer design programs, and generally they cost around 3$ for a tube when you design and submit them to, say, IDT. Easy... though depending on what you want to do, you may need to buy large numbers of them ^.^ Especially if you want to work with the genes of some non-fungal eukaryotes, many of whom have tons of introns. Basically, primers are small price apiece but can be needed in great abundance. Even if you use published primers so don't have to design your own, you will need primers.

Electrophoresis setup: This will be used all the time. I made mine, and it works well enough, though I bought the power supply for for 60$ at surplus because I didn't feel like trying to build my own (the high current diodes are expensive, and I just wanted to get started at that point).

heat blocks (with the blocks for tubes): For many purposes, there will be incubations at, say, 90*c, of 65*c, ect. These can be pretty cheap used, I have several that I use for different temperatures and I don't think I've spent more than 200$ on all of them... but getting the right blocks can be a little difficult, one of mine I bought for close to 100$ simply because it came with four blocks that I needed (though it works fine as well).

Pipettes: You will need a set of pipettes, and it will need to span from 1ul (or less) up to 100ul at least (1000 better). These can be on the pricey side: I got mine for ~70$/pipette, but they were calibrated and everything, so my whole set spanning 0.1ul to 1000ul cost ~350 (small so shipping negligible). What can I say: I splurged.

Freezers: I've never needed a -80*c freezer: almost anything can be done and stored with a -20*c freezer, and I got mine at the local farmers supply store. It's a little things that works well for my purposes. I'de like to get a small -40*c as well, which are good for long term storage but don't need Liquid Nitrogen to maintain, but that's on my wish list.

Reagents other than TaqPol: There's a bunch of other reagents that one may need. These can add up. Kits for DNA extraction (these can get a bit up there in price per reaction, though there's methods published for free online for the experienced... but if starting out, go with a kit.), dNTPs, nuclease free water, PCR tubes, enzymes like restriction endonucleases and DNA ligase (depending on what you are doing), pipette tips, agarose (for gels), Big Dye if you want to sequence things, 100% ispropanol, nondenatured ethanol, and MANY others. You'll have to figure out what you need from whatever protocol you decide to go with. Some places like universities can get discounts from some companies like Fisher and Sigma, and I have been able to cut my costs on these by going through my university (and reinbursing them out of pocket).

Sterile hood: This may or may not be applicable to a given person, since most PCR can be done on a benchtop, but for innoculations you want sterile conditions, and if working with fungi some of the major contaminats will often amplify and screw up your stuff. I wanted one fireproof si I could flame sterilize things, so I built mine out of sheet metal, corner bead, and bolts. It took a month, cost about 200$, but it is a nice internal flow system. My friend built a sealed system out of a 7$ clear plastic container, a pair of kitchen gloves, some duct tape, a thing of silicone caulk, a cheap household fan, and a small HEPA filter, total cost 50$ and one day to assemble. You WILL want a HEPA filter no matter what you do: that's essentially what professional biohazard hoods use (they are tested at the factory: any that meet biohazard specs go to biohazard hoods, everything else goes to stores for everyone else... but the store ones still work wonderfully. As long as you keep the flow internal (not bringing in outside air), the filter should last just about forever.

Pressure cooker: For 'autoclaving', you will want a pressure cooker to sterilize things and make sterile media. I have two, one large and one small, and they cost 60$ and 100$ respectively if my memory serves. I also do canning so they are multi-purpose and well worth the cost.

Sequencing: If you ever want to do sequencing, you will need to 'use' a sequencer. Sequencers CAN run no less than tens of thousands of dollars, and usually hundreds. However, if you need the capacity of your own sequencer, you are already a high throughput lab and can likely afford it, because often they can take a 96 well plate at a time, and do 20some plates per day, so we're talking thousands of amplicons daily... and they last a good long time, too, even under high use situations. DON'T buy a sequencer. Send your sequenced samples off to the nearest place with one. Universities often charge no more than a few dollars per sample to run your sequence for you, and in doing so cover the costs of maintenance and parts. This can certainly add up over time, but is rarely a large cost at any single time, so just send those samples off and wait a few days for the sequence to come back.

 

The gist of all this is that, if you take your time, be frugal, buy used, learn some electronics, and are willing to have a few failures, you can start doing basic genetic techniques for not a six figure sum, but as low as the lower four figures (with increase over time as you use more and more reagents, but that's inevitable). It's still not necessarily cheap, but it depends on your interests, and there are many people with more expensive hobbies (cars, pro bicycles, landscaping, photography, boating, travel, and many more), and it can be very rewarding and happymaking (not to say the others on the list are not rewarding and happymaking as well). That said, however, I've yet to sucessfully modify anything, and am at a standstill right now due to having too many other things to do, but it's all part of the trial and error, and when I have more time than I currently do I will actually get back to working on things.

[Arete]

That said, the field is nowhere near as expensive to get into as the above says... you can start doing basic genetic techniques for not a six figure sum, but as low as the lower four figures

 

 

Ultimately the OP didn't want to do basic PCR and vector cloning, but actually wanted to produce mutant plants and vertebrates, so we're comparing apples and oranges, and actually, what I specifically said was: "setting up Sanger sequencing will set you back a decent six figure sum".

 

You outsource your Sanger sequencing to a core facility. I can see that as a major self sufficency hurdle for a hobbyist. We just had the capillaries replaced in our aging 3730xL and it cost around $7k, so both the purchase and the upkeep will be expensive. Even if you buy an old ABI 377 slab gel sequencer, you'll have the issue of trying to safely use large amounts of acrylamide at home.

 

I'm glad you managed to get a home setup running as cheaply as you could - you could go even cheaper: instead of using a thermocycler you could go competely old school and set up three water baths, transferring your pcr reactions to each in sequence One issue I can see is that some of your equipment would fall outside of our facilities' health and safety parameters (e.g. home made laminar flow hood), and some would fall outside of the standard operating procedures required to publish results by our funding body (e.g. cloning vectors stored above -80C).

 

Our lab recently bought an Ion Torrent next gen platform which - for a sequencer capable of high throughput genomic sequencing was pretty cheap ~ $80,000USD. That's not that far out of reach for a hobbyist I imagine. Again, the service protocol would be prohibitively expensive, however and you need to have a pretty high throughput of samples to keep the machine in service. http://seqanswers.com/forums/showthread.php?t=11929

[CharonY]

three water baths, transferring your pcr reactions to each in sequence

 

A classic setup was three oil baths and an undergrad.