You will be given stock tubes containing DNA in the following concentrations: Cu

Question

You will be given stock tubes containing DNA in the following concentrations: Cut pET41a(+) DNA: 25 ng/ul egfp insert DNA: 7 ng/ul Uncut pET-41a(+)/EGFP recombinant plasmid DNA: 25 ng/ul Ligation #1: 1:1 molar ratio pET-41a(+) vector: egfp insert 50 ng NcoI/NotI cut pET-41a(+) DNA_____ul 7 ng egfp insert DNA______mu 1 sterile dH2O_____pi 10x ligase buffer_____ul (final concentration will be 1X) DNA ligase: 1 ul Ligation #2: 1:3 molar ratio pET-41a(+) : egfp insert 50 ng Nco/NotI cut pET41a(+) DNA_______u1 21 ng egfp insert DNA_____mu 1 sterile dH2O_____mu 1 10x ligase buffer_____mu 1 DNA ligase 1 mu 1 "Ligation" #3 (positive control): circular (uncut) recombinant pET41a(+)/EGFP plasmid (why?) 50 ng uncut pET-41a(+)/EGFP recombinant plasmid DNA _____mu 1 sterile dH20_____mu 1 "Ligation" #4 (negative control #1): No DNA or DNA ligase control (why?) sterile H2O_____mu 1 "Ligation" #5 (negative control #2) No DNA ligase control (why?) 50 ng Nco/Notl cut pET41a(+) DNA_____ul 21 ng egfp insert DNA _____mu 1 sterile dH2O_____mu 1 10x ligase buffer_____mu 1

Explanation / Answer

C1=25

C2=50

V1=1

V2=x

C1V1=C2V2

V2=2

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