Week 2--"Take 2"

Last week Melissa was working to build her library of mutant monomer Cyanoviran protein. She put us right to work in the lab which was really exciting for me. We made phage, we mixed LB broth and agarose, added enzymes to DNA, combined E.coli with media and while doing all this became familiar with how to use the autoclave and pipettes.

We had a little trouble with accuracy in pipetting, so we practiced a lot! Pipetting needs to be precise. It involves first calculating how much you need and then setting the pipette to the proper setting for the prescribed amount. (This can be tricky if you happen to pick one up that is "missing the last zero" on the setting indicator! And this is where we use math in the lab as well as the metric system--the ability to convert from micro to milli (and vice-versa)quickly is helpful! Drawing the fluid into the pipette is fairly easy; you just push the button down (only to the first "catch") before dipping into the solution. Then you release the button while holding the pipette steady. Next you release the fluid from the pipette into the new container. Depending on the container and the substances you are working with this can be tricky--I was having trouble because I kept getting air bubbles in the pippette and sometimes not able to release the total amount. Another problem was that I kept shaking upon the release when I was realising into a test tube. This is not a problem with just one solution but if you have multiple solutions you risk contaminating the container of original solutions. When you do this you have to discard the pipette tip and get another one.

Friday was supposed be a big lab day; however, before we arrived at 8:00 a.m. Melissa realized that the lack of "clear spots" in the petri dishes we had prepared Wednesday afternoon was due to a missed step in the process. She had gone back to her manual and re-read the protocol finding that she had not put in an enzyme to dissolve the binding on the DNA used for building the library. So we started the process again this week.

It is now Thursday morning. Melissa is again baffled because there are even fewer clears spots (indicating phage) in the petri dishes.


I have a new digital camera and have been collecting photos on it and my phone...I'm going to become a real techy this weekend and learn how to upload phone and cameras to my computer and then post them here.