Wednesday, August 11, 2010

A day in the life of Liz (at duquense)

Every day Liz is either doing transformations, mutatations, or somedays, both. Today she decided to do a transformation for us. She begins in this first picture by preparing the auto-claved pipette with a sterile tip before taking 1 µL of plasmid from a tiny test tube.

Then the plasmid is added into a sterile 14 mL polypropylene tube (which must be used because it is VERY sterile), while making sure not touch the sides of the tube with anything but the tip of the pipette because that is the only sterile part of the apparatus.

The last step before heat-shocking the sample is to add 50 µL of XL-1 blue Supercompetent cells to the polyproylene tube which will later absorb the plasmid into the cell, thus transforming it. After that the final transformations are sent to Pitt for sequencing, which usually takes about a week to get results. Most of the time the mutations are unsuccessful because of unknown reasons that could include an instrument or apparatus being unsterile, or a problem with the DPN1 restriction enzyme .

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