So qPCR is still kicking my ass. Integrated DNA Technology actually tweeted me back in response to my moaning about LNA oligos, which made me smile even if it wasn’t especially helpful.
Essentially: my primers may be awesome (provided I can acquire an unrealistic amount of DNA) or they may be unusable. I can’t tell without testing them with a stupid amount of cDNA. (Which I don’t understand, because they worked in regular PCR just fine).
Option A) I admit that qPCR with LNAs is not going to work and go back to hunting for a new solution
Option B) I spend today extracting RNA from some leaf tissue I have, which I was saving, but almost certainly don’t need, so I can make a shedload of cDNA for the purposes of testing them.
Option C) I sit and cry.
Basically, when stuff goes wrong in my PhD I like to sit and consider my options for a long time. This is strange, because in real life I think fast and trust my gut instinct. Unfortunately, this hasn’t stood me in good stead in science, so now I tend to over think.
Conference is in just over a month, but the uni is closed for a long time over Christmas. So I need to make a decision now. Engage brain! Engage!
(Today is all about tom yum ramen. I’m going to have to start eating bananas to counteract all the sodium in these things…)