{"id":734,"date":"2009-08-19T22:18:38","date_gmt":"2009-08-19T22:18:38","guid":{"rendered":"http:\/\/occamstypewriter.org\/mindthegap\/2009\/08\/19\/in_which_i_get_what_i_pay_for\/"},"modified":"2009-08-19T22:18:38","modified_gmt":"2009-08-19T22:18:38","slug":"in_which_i_get_what_i_pay_for","status":"publish","type":"post","link":"https:\/\/occamstypewriter.org\/mindthegap\/2009\/08\/19\/in_which_i_get_what_i_pay_for\/","title":{"rendered":"In which I get what I pay for"},"content":{"rendered":"

Sometimes it feels a little bit too<\/em> good to spend money in the lab.<\/p>\n

\n\"\"
\nBlue streak<\/strong> If you have to ask, you can’t afford it<\/em><\/p>\n

\nFor those of you following the story so far, I recently triumphed in my new career as a Cloner For Hire<\/a>. In brief, I managed to make and verify three complex DNA constructs, finishing the very day my visa-frazzled colleague returned to London to pack for his sabbatical. This had already turned out to be a bit more involved than I had initially anticipated \u2013 and then the boss suggested I extend the favor by recombining the little blighters into baculovirus vectors (so-called ‘bacmids’) so that my colleague could furiously finish up a few final experiments for the paper in the week before his departure.<\/p>\n

\nThe bacmid kit<\/a> seemed straightforward, but I did notice that the transformation step \u2013 the part of the procedure when you zap bacterial plasmids into E. coli<\/em> and allow them to recombine with a resident bit of DNA \u2013 looked a bit problematic. I could tell something was up by the cagy way that the kit’s instruction manual phrased certain things. That the procedure was far from efficient was also clear by the modifications in the standard protocol: five hours of growth before plating onto the triple antibiotics plates instead of thirty minutes; two days’ growth after plating instead of one; and then a further re-streak of ten<\/em> prospective white colonies to confirm that they actually were bona fide recombinants. <\/p>\n

\nAt first I thought the company was just trying it on when they suggested purchasing their own special \u2013 and highly expensive \u2013 derivative of X-gal, the chemical substrate you need to put into your agar plates so that you can tell which bacterial colonies have successfully recombined (blue colonies are unperturbed, whereas white colonies are bacteria that have lost the beta-galactosidase gene because the genetic modification of interest has disrupted it). But then I thought about it: two days<\/em> to get color development and a re-streak to confirm it? These colonies were likely to be pretty anemic, and even freshly prepared X-gal can often result in very pale colonies. We simply didn’t have a day to lose on ambiguity. <\/p>\n

\nDear reader, I bought that expensive X-gal derivative. And I have never seen such lovely, deep-blue, almost slate-purple colonies in my life. I thought I would weep with joy, and my colleague as well \u2013 it was KimWipes all around. And it’s a good thing I did too, because after re-streaking the white colonies, the blue controls were so faint the next morning that I spent many, many minutes squinting at the colonies, trying to decide if there was the faintest whiff of blue about their whiteness.<\/p>\n

\nThe moment of truth came in the form of PCR confirmation. There is a wonderful moment in science just before a result, no matter how routine. For me, even after two decades in the lab, I still get a frisson of anticipation when an agarose gel is sitting on the lightbox, the verdict unclear in that hushed moment just before the UV source is switched on:<\/p>\n

\n\"\"<\/p>\n

\nAnd then \u2013 <\/p>\n

\n\"\"<\/p>\n

\n\u2013 you see that your bands of DNA spell out success. It’s a priceless feeling.<\/p>\n

\nJust an hour before the Fedex deadline, I was coaxing my final bacmids into solution and labeling up the tubes to ship them off to my colleague’s sabbatical lab; with a prevailing wind, they’ll rock up at about the same time tomorrow morning as he does.<\/p>\n

\nTime, I think, to saddle up and head off into the sunset.<\/p>\n","protected":false},"excerpt":{"rendered":"

Sometimes it feels a little bit too good to spend money in the lab. Blue streak If you have to ask, you can’t afford it For those of you following the story so far, I recently triumphed in my new … Continue reading →<\/span><\/a><\/p>\n","protected":false},"author":6,"featured_media":0,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1],"tags":[],"class_list":["post-734","post","type-post","status-publish","format-standard","hentry","category-uncategorized"],"_links":{"self":[{"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/posts\/734","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/users\/6"}],"replies":[{"embeddable":true,"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/comments?post=734"}],"version-history":[{"count":0,"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/posts\/734\/revisions"}],"wp:attachment":[{"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/media?parent=734"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/categories?post=734"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/occamstypewriter.org\/mindthegap\/wp-json\/wp\/v2\/tags?post=734"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}