Objective: To veriffy the presence of the LuxOD47E gene in the psB1AC3 BBK vector
Size of plasmid Backbone: 3055 bp
Size of the gene of interest: 1362 bp
Total size: 4417 bp
Sequencing needs 100ng of DNA/ 1 kb
4.4kb x 100ng/kb = 440ng
Concentration of C2= 92.9ng/uL
440ng/ 92.9 ng/uL = 4.8 uL DNA
Diluted 4.8 uL DNA (LuxOD47E BBK C2) with 5.2 uL ddH20 and sent down to Univeristy of Calgary Sequencing Labs with VF2 and VF1 primers.
Results: Sequencing came back and results indicated that the gene if interest, LuxOD47E was not in the psB1AC3 vector. We will have to go back to the gene in the pCR2.1-TOPO vector and try to get it into the psB1AC3 vector.
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