Sbb14-jpn: Difference between revisions
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--[[User:Joshua P Nixon|Joshua P Nixon]] 16:15, 14 February 2014 (EST) | Today we created a [https://docs.google.com/spreadsheet/ccc?key=0AsFOPcCYYB1DdFJ4TmwzdTY4ZngzYjZ4NEY4eFIwS3c&usp=sharing design spreadsheet]which we will use to design oligos for cloning our assigned aminoacyl tRNA synthetases--[[User:Joshua P Nixon|Joshua P Nixon]] 16:15, 14 February 2014 (EST) | ||
We determined that because alanyl-tRNA synthetase is a homodimer that the other related sequence we had found (375 aa) is not another chain to the protein and unnecessary for our experiment. We also made a choice between the two aspartyl-tRNA synthetases for the one which is not of the nondescriminating variety. | |||
--[[User:Joshua P Nixon|Joshua P Nixon]] 13:30, 18 February 2014 (EST) | |||
Today we started the wetlab portion of our experiment by miniprepping the Pbad-Oled from E. Coli so that we may later clone our aminoacyl tRNA synthetase constructs into it (We use a Quiagen miniprep kit). --[[User:Joshua P Nixon|Joshua P Nixon]] 01:19, 28 February 2014 (EST) | |||
Somewhere in here we ran PCR's and gels. | |||
Today we started did digestions for all but alanyl-tRNA synthetase, we also designed oligos to amplify the context sequence around alanyl-tRNA synthetase and isoleucyl-tRNA synthetase. This is because for A we were getting two bands with the one of incorrect length being longer and for I we had a very weak band, after doing this context PCR we will try to PCR the genes off that product. Also we will try to gel purify out the weaker band for A. --[[User:Joshua P Nixon|Joshua P Nixon]] 13:45, 8 April 2014 (EDT) | |||
Latest revision as of 10:45, 8 April 2014
Today we created a design spreadsheetwhich we will use to design oligos for cloning our assigned aminoacyl tRNA synthetases--Joshua P Nixon 16:15, 14 February 2014 (EST)
We determined that because alanyl-tRNA synthetase is a homodimer that the other related sequence we had found (375 aa) is not another chain to the protein and unnecessary for our experiment. We also made a choice between the two aspartyl-tRNA synthetases for the one which is not of the nondescriminating variety. --Joshua P Nixon 13:30, 18 February 2014 (EST)
Today we started the wetlab portion of our experiment by miniprepping the Pbad-Oled from E. Coli so that we may later clone our aminoacyl tRNA synthetase constructs into it (We use a Quiagen miniprep kit). --Joshua P Nixon 01:19, 28 February 2014 (EST)
Somewhere in here we ran PCR's and gels.
Today we started did digestions for all but alanyl-tRNA synthetase, we also designed oligos to amplify the context sequence around alanyl-tRNA synthetase and isoleucyl-tRNA synthetase. This is because for A we were getting two bands with the one of incorrect length being longer and for I we had a very weak band, after doing this context PCR we will try to PCR the genes off that product. Also we will try to gel purify out the weaker band for A. --Joshua P Nixon 13:45, 8 April 2014 (EDT)
