... is with me.
Two constructs to make.
Two colonies I had.
Each trial a single clone.
I cut them, they cut just the way I hoped.
Each clone was sequenced.
The sequence told me that I have what I need.
Although my hopes were low, I did succeed.
And now the clones are sent into the fire ...
Will they do my bidding?
Will they tell me the truth?
(I feel like Nick Cave)
PS Cloning = make an exact copy, in this case a DNA construct. Actually what I did was construct a a gene on a DNA vector and then introduce the newly made reagent into bacteria so that the cells would copy the novel DNA fragment. I would never clone a human. Promise.
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Just a single construct to make.
Hundreds of colonies I had.
Each trial 36 clones.
I cut them, they just didn't cut the way I hoped.
No clone was sequenced.
The sequence would have veryfied that I have not what I need.
Although my hopes were high, I did not succeed.
Never mistake PvuI for PvuII, especially if a BamHI/PvuI and a BamHI/PvuII fragment have the same size and you fill in the ends for blunt end ligation.
And rather do refer to your lab book immediately where it is documented that you have used the wrong enzyme instead of waiting a week before looking it up (due to bad experiences my documentation really improved since my time as a PhD student).
To quote Nick himself:
The ladders of life we scale merrily
Move mysteriously around
So that when you think you're climbing up, man
In fact you're climbing down