User:Howard Boland/Notebook/Art from Synthetic Biology/2010/09/16
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Based on the confirmed digest -link needed- I did a nanodrop of the 3 samples in preparation for sequencing.
In order to prepare my samples for sequencing the following concentrations were prepared
Dilution calculation: 48nmol in 480nl of RNAse free water gives 0.1nmol/ul = 100 pmol/ul thus to obtain the concentration for the sequencing we will need 6ul x 4pmol/ul) / 100pmol/ul = 0.24ul of primer in 5.76ul of water (total 6ul)
I prepared both my primers as follows: μL
I prepared my vector by taking 20ul:
The order sheets and specifications where attached with the samples and sent to the Wolfson institute for sequencing.
End Overnight Growth
I took at the two samples from the overnight growth
I set up a miniprep using one of today's overnight growth and one from the four prepared yesterday. The samples extracted using QIAgen Miniprepp kit and eluted in 30μL water.
The extracted plasmids (two samples) from the miniprep were each immediatly digested using
The digestions where run on a 2% Agarose Gel