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Daily Lab Notebook

1) Amplification of pvdQ gene using PCR from Pseudomonas aeuriginosa genomic DNA. The designed primers contain the prefix and suffix so that typical digestion for insertion into iGEM’s standard vector can be subsequently performed.


2)Test digestion of amplified pvdQ with EcoRI only, EcoRI & SpeI, PstI, and EcoRI & XbaI.

3) Repeat the digestion of pvdQ with EcoRI and SpeI multiple times to obtain the desirable conditions of the digestion to prevent Star Activity.

4)Mass single digestion of the amplified pvdQ with EcoRI. Desirable fragment size should be about 5,600bp. The fragments that result due to Star activity are 3,800 and 1,800bp.

5)Gel purification and Nano-drop concentration check of digested pvdQ.