November+2011

=toc November 2011=

Week of October 31
**October 31, 2011** Today we did a mini-prep of our 8 different culture tubes. We also decided on which primers to use for cutting our vector. We also looked at the gel from our single digest and we concluded that Ecor1 didn't cut. //**Mini-Prep **//
 * 1) Dump out the medium -- pellet in the bottom
 * 2) Add 250 microliters r3 buffer which is used to resuspend the cells
 * 3) Transfer to a 1 mililier tube
 * 4) Votrex for 30 seconds
 * 5) Add 250 microliters L7 -- invert tube 10 times
 * 6) Wait 5 minutes
 * 7) Add 350 microliters M -- invert 10 times -- this will form a percipitate
 * 8) Centrifuge for 10 minutes -- clear solution will stick to the column
 * 9) Centrifuge for 1 minute -- add 500 microliters W10 (ethonal)
 * 10) Centrifuge for 1 minute 00 200 microliters of W9 (ethonal)
 * 11) Centrifuge for 1 minute

media type="custom" key="11723118"
 * November 2, 2011**

Today we looked at our gel using Ecor1, BamH1, and Sma1 as shown above ^. And we also went through our vector map in detail: media type="custom" key="11723136"

Week of the 7
**November 7 & 9, 2011** Today we learned how to make our own primers for our pGlo GFP sequence. We took the following notes: media type="custom" key="11723316" And this was the gel of our digest from monday: media type="custom" key="11723344" You can clearly see the two distinct bands in our gel

Week pf the 14
Today we ran our gel of our GFP and the vector which separated into two linear bands. We then cut them and dissolved them from the gel into pure DNA and vector. media type="custom" key="11723528" media type="custom" key="11723552" These were the instructions for dissoliving the vector and pGlo GFP DNA from the aragose gel^ media type="custom" key="11723504" And these were our final notes on how to create our own forward and reverse primers of Sma1 and Nhe1^
 * November 14, 2011**

Week of the 21
Today we performed transformation our bacteria.
 * 1) E. coli bacteria should remain cold
 * 2) add 40.0 microliters into microcentrifuge tube
 * 3) let it sit on ice for 2-3 minutes
 * 4) add 10.0 microliters of ligation reaction to the bacteria
 * 5) mix and then put back on ice for 20 minutes
 * 6) after 20 minutes, put on 42 degree waterbath for 60 seconds
 * 7) add 500 microliters of SOCS broth
 * 8) shake overnight at 37 degrees

Week of the 28

 * November 28, 2011**

Francis showed me how to create ampicillan plates that we usually use for our bacteria.
 * November 30, 2011**