I took a couple of days last week to finish the Merlin cloning experiement but to no avail. I didn't get any colonies on my LB/amp plates. Not even in the positive controls. I decided to check my vector to make sure it was being digested properly. Yesterday, I did 8 restriction digests in an attempt to figure this out.
This project is my Moby Dick. I have been working on it for so long that I just can't conceive of not finishing it. It has been a real source of irritation. It is a very straight forward cloning experiment that should go off without a hitch - even with the variables that are inherent to biology. I think I have narrowed down the problem to the ligation reaction. I redid the experiment today so I guess I'll see if I solved the problem tomorrow morning if I have colonies on my plates.
My boss has been out of town for the last two weeks and the bulb housing for the mercury bulb in our epifluorescent microscope burned out. This means that all the immunofluorescence experiements for the last week have been put on hold. Unfortunately, nobody told the cells to stop growing and we have had to come up with experiments using assays other than immunofluorescence. Normally this wouldn't be much of a problem but I am not up to date with everybody's progress and therefore don't necessarily know what needs to be done or redone. It has been a real pain in the neck. Marlan is back tomorrow and then starting Friday, I will be out for a week and a half. The break will be nice.
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