finally! for the love of god, it finally worked. i ran a PCR screen where i get a 1600 bp band for an ‘insert containing’ clone (ie, what i want!) and a 1200 bp band for any negatives. and i got . . . like 20 positives!! my negative controls worked, too, so i’m not just wishfully pretending to read the ladder wrong, or something. i almost posted a picture of the gel (it was a rather attractive one, too, a double-decker with 20 lanes on each tier for 40 total!), but i realized that it would bring my blog up to a new level of dorkiness that i never really want to reach.
i have to send the clones that were positive by gel on to sequencing, since each clone only has a 50% chance of having the insert in the correct orientation. sequencing will also make sure there were no spontaneous mutations that would render the whole thing useless.
sure hope that doesn’t happen.
anyway, i’m very happy. i am going to treat myself to a new j.crew bathing suit as a reward. see, i’m not that dorky.
ps: if you looked at the bathing suit link, don’t worry – i am NOT getting whatever heinous color combination they have there, it just comes up as the default. i’m going for the rather demure chicory brown color, because i am in that sort of mood. though the paisley print is sort of cool, too.