The days are still going by like minutes...
Since Monday, I've begun preparations for the next (albeit unofficial) stage of my project, but I'll get to that. On Tuesday, I was the only person in the lab, since Suzanne (the research specialist) was out, and none of the undergraduate workers or volunteers were available. ULTIMATE POWER...means nothing when you're the only one in the wasp lab. So, that went well, since all I had to was a little bit of culturing for about three hours. I actually had only about 80 minutes-worth of work to do, but I was able to find a secret Texas Encarsia pergandiella harvest that wasn't on the schedule. As it turns out, two weeks ago, that culture was accidentally infested, and we've just been rolling with it ever since. I'd just like to say right now that although this isn't a problem, the E. pergandiella wasps are not my favorites to harvest because their pupae are just so unappealing to look at. They're the scrawniest, palest pupae in the lab. Well, they're still easier to tell apart from whitefly pupae than are Eretmocerus pupae.
On Wednesday, the most important stuff occurred!!! I isolated about 200 pupae from 2 different Eretmocerus emiratus cultures that we're going to compare on Monday during our special mating experiments. The two different cultures we're going to be using are the Rickettsia-positive/Wolbachia-positive E. emiratus and the Rickettsia-positive/Wolbachia-negative E. emiratus. Unlike the wasps we used for my experiment, these ones are R+, instead of R- (this is a result of raising these wasps on R+ whiteflies; Rickettsia is horizontally transmitted), but this shouldn't really matter all that much, since Rickettsia hasn't been shown to do anything to E. emiratus. The experiment, of course, will determine whether a female wasp has a preference for a male wasp of a certain infection status. We will be using both W+ and W- females, and seeing whether they can tell that a male is W+ or W-, if at all. Of course, the W+ females act as more of a control, as do the W- males, since the interesting cross is between the W- females and the W+ males, because the W+ males can sabotage the reproduction of the W- females with their unusable, infected sperm. The W- female could still produce males from this cross, since they develop from unfertilized eggs, as is the case with many hymenopterans.
On Thursday, the most consequential thing that I did in the lab was assisting with another PCR to determine the effectiveness of various PCR primers. This time, we tried the primer CO1 (which worked kind of well before), but with twice the amount of magnesium chloride, and alongside this, we tried EF (elongation factor) made from Encarsia pergandiella proteins. We didn't use BSA (bovine serum albumin) on the CO1 PCR and one of the EF PCRs. In this case, the BSA, which we used on the other EF PCR, did not have a significant effect. The CO1 PCR with double magnesium chloride, worked very well.
Today was a relatively standard day... most of what I did was culturing, alongside the undergrads. There was a birthday, though, so there was lemon cake and mini chocolate muffins, but I didn't have any. Oh yeah, those pupae I isolated on Wednesday have begun to emerge, but the vast majority are still developing. The sex ratio doesn't look too biased...
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