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.