Difference between revisions of "Romina Clemente Notebook"
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== [[Roclemente]] 9:23, 3 June 2009 (EDT)== | == [[Roclemente]] 9:23, 3 June 2009 (EDT)== | ||
Revision as of 13:46, 3 June 2009
Roclemente 9:23, 3 June 2009 (EDT)
Shamita and I are trying to find suitable reporter proteins to use. Yesterday, Leland and Alyndria were working on ways to insert the gene sequences into the plasmid. Upon seeing how they wanted to manipulated the reporter gene to include the logical clauses, we came up with a few criteria for the reporter genes we would use. The following criteria for genes are listed in order of the broadest aspect to look at to the narrowest aspect:
a) Doesn't contain restriction sites for the 4 restriction enzymes (EcoR1, Xbal, Spel, Pst1) used to cleave the Biobrick part out of the plasmid.
b) Contains 6 cutter restriction sites.
c) These restriction enzymes aren't blunt (cleave straight down at one spot).
d) These restriction sites are close to thge 5' (beginning) end of the sequence.
e) These enzymes are easiest to work with and cheapest.
We are finding the part numbers of the reporter genes we want to use (antibiotic resistance, fluorescence, LacZ) through our own GCAT because we know these ones work. We are then locating these parts on the parts registry [1] website. We copied and pasted the gene sequences we obtained from the registry onto the ApE software [2]. From here, we were able to generate a genetic map of each gene that outlined each restriction site that fit our criteria. We put each genetic map, alongside the part number used, into a Word document.
