As I stated in my last post of this nature, my goal for this week was to combine all three PCR products and get them ligated into the first vector, pGEMTeasy. In short, I failed. However, I did get quite a bit accomplished. For ease, I've posted the picture from last week. (Come on, you know you love it.)
As far as the PCR goes, I completed PCR round three for 20 of the 26.
Of those 20, 13 of the PCR products were successfully ligated into the first cloning vector and are getting sent off for sequencing today. (I am dreading looking at all this sequencing data. I freaking hate it! It bores me to tears.) I will ligate the remaining 7 PCR products (which I just obtained overnight) over the weekend and transform them on Monday.
So, in all, not quite where I wanted to be, but not exactly too far behind either.
My goal for the end of next week is to get the final 6 PCR products created (i.e. combine segments 1,2 and 3) and ligated into the first cloning vector. Obviously, everything needs to get sequenced. Finally, I hope to sub-clone at least half (that's 13) of the pGEMTeasy constructs into the final vector.
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5 comments:
you have succeeded in pcring (if that is a word) more fragments and sequencing them than I have.... I should get mine going. It's just that the specific primers have ended up with several bands....
strange how that happens sometimes, right?! ;) TGIF! For non preggers that means drinks :)
"am dreading looking at all this sequencing data. I freaking hate it! It bores me to tears"
flashback, FLASHBACK!!!!
I did loads of this kind of thing during my PhD and postdoc (making promoter mutants, mostly) and you're right, it's a drag. Good music and coffee both help. Good luck with the remaining cloning!
Glad to materialize here. Good day or night everybody!
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What do you use to view your sequencing data?
Oooh, I so wished for some coffee in those last weeks, but I never did come up with a better pick me up for those dreaded tasks. Good luck!
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