…by which I mean, it’s not always the best tool for the job, but everyone insists on always using it anyway, and has a variety of excuses for doing so…<\/p>\n
Honestly – I’m trying to set up a clone library of 16s rDNA sequences using this kit. Never mind which kit it is – it actually does seem to work. I was just struck by the amount of hassle involved in shipping and storing the kit and it’s supply of “competent<\/span> cells”.<\/p>\n When you get them, take them out of the dry-ice they’re shipped in and put them in the -80?C freezer immediately<\/em> or they’ll die! Only thaw them carefully just before you use them, and do it on ice or they’ll die! Don’t heat-shock<\/span> them for more than exactly 30 seconds or they’ll die! Once you’ve got them growing, you have to keep moving them to fresh selective<\/span> media frequently or they’ll die! Or, you can carefully place them in the -80?C freezer…or they’ll die! Don’t look directly at them or they’ll die! (Do Not Taunt HappyFunCell!…)…<\/p>\n Seriously, running those gigantic -80? freezers can’t be cheap. Wouldn’t it be more convenient if you could grow up your transformant<\/span> as an ordinary culture and just add your DNA samples and some kind of inducer chemical to make them take it up? Surely there must be some other organism that might be made to work like that.<\/p>\n Actually, it seems a number of the “Gram-positive” (firmicutes) organisms can enter a state of “natural competence”, where they naturally take up double-stranded DNA molecules from the environment. Bacillus subtilis<\/em> is one. I’ve even seen references to “natural-competence” based protocols for transforming B.subtilis<\/em> (or other Bacillus<\/em> species, presumably) but it only seems to be in an out-of-print, $400 book.<\/p>\n Wouldn’t that be more convenient (using B.subtilis<\/em> that is, not the $400 book)? Plus, when you wanted to store your transformed culture for later use, you could just heat the culture up to, what, about 55?C for 15 minutes or so (as I recall) then let it dry. The spores will contain whatever “bonus” plasmid DNA you added (if spores didn’t keep plasmids, then anthrax wouldn’t be such a danger…) and will last practically forever<\/span> at room temperature. Mix the spores with some dried nutrient powder and seal them in a foil packet. Instant transformants, just add water!<\/p>\n But NOOOOO…..”But, everybody else<\/em> uses E.coli, so I have to.” “They only make ‘BogoGen SuperMiniUltraKlone Kit 2000’ with E.coli, and we have<\/em> to use that!” “But, nobody knows that other stuff, but everybody’s already familiar with E.coli!” “I’m a BogoGen Certified E.Coli Engineer, and I say everything else is just a toy and doesn’t work!” “All the books and stuff are about E.coli…”<\/p>\n Bah! Pathetic excuses. Anybody got a huge wad of venture capital to throw at me? The more I think about this, the more I think ‘untapped niche’…Heck, the electricity savings on not having to run a -80?C freezer constantly alone ought to qualify for a good “Fight Global Warming – Say ‘No!’ to E.coli!” marketing campaign…<\/p>\n Bonus perk: All the natto<\/span><\/em> you can eat…<\/p>\n","protected":false},"excerpt":{"rendered":" …by which I mean, it’s not always the best tool for the job, but everyone insists on always using it anyway, and has a variety of excuses for doing so… Honestly – I’m trying to set up a clone library of 16s rDNA sequences using this kit. Never mind which kit it is – it … Continue reading E.coli – the “Microsoft” of the biotech world?<\/span><\/a><\/p>\n","protected":false},"author":1,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[4,15,6,5,11,8],"tags":[],"class_list":["post-47","post","type-post","status-publish","format-standard","hentry","category-feedback-solicitation","category-methods","category-microbiology","category-nerdity","category-rant","category-why-does-it-work"],"_links":{"self":[{"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=\/wp\/v2\/posts\/47","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=\/wp\/v2\/posts"}],"about":[{"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=%2Fwp%2Fv2%2Fcomments&post=47"}],"version-history":[{"count":0,"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=\/wp\/v2\/posts\/47\/revisions"}],"wp:attachment":[{"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=%2Fwp%2Fv2%2Fmedia&parent=47"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=%2Fwp%2Fv2%2Fcategories&post=47"},{"taxonomy":"post_tag","embeddable":true,"href":"http:\/\/www.bigroom.org\/wordpress\/index.php?rest_route=%2Fwp%2Fv2%2Ftags&post=47"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}