a b What physical forms could you expect in the pGLO plasmid

a)

b) What physical forms could you expect in the pGLO plasmid?

c) Should the number of different DNA bands in the \"double digest\" lane equal the number of bands in the EcoRI pGLO digest and NdeI pGLO digest combined? If so, does this make sense? Why?

SAMPLE	PREDICTED SIZE OF FRAGMENTS
P-pGLO DNA uncut
E- EcoRI pGLO digest
N- Ndel pGLO digest
D- EcoEI and Ndel pGLO double digest

5.0 Kb araC BstE 11 BamH .Nde ! Nhe I Hind III Nde I bla BamH I 3.0 Kb EcoR Ps m ee ha 1.0 kb ind 111 > y023 I o13 Nde ¾ !JO 4.0 kb

Solution

a)

SAMPLE

PREDICTED SIZE OF FRAGMENTS

P-pGLO DNA uncut

5 kb

E- EcoRI pGLO digest

5 kb

N- Ndel pGLO digest

3.3kb, 1.8kb and 0.2kb

D- EcoEI and Ndel pGLO double digest

2.8 kb, 1.8kb, 0.5kb and 0.2kb

b) What physical forms could you expect in the pGLO plasmid?

pGLO plasmid will is found in circular form

c) Should the number of different DNA bands in the \"double digest\" lane equal the number of bands in the EcoRI pGLO digest and NdeI pGLO digest combined? If so, does this make sense? Why?

Yes. As we can see from the vector map that EcoRI digest the Plasmid at the site which is between the two NdeI enzyme whose fragment size is 3.3 kb. Upon double digestion, we got two bands of 2.8 kb and 0.5kb which upon adding make the fragment of 3.3 kb

SAMPLE	PREDICTED SIZE OF FRAGMENTS
P-pGLO DNA uncut	5 kb
E- EcoRI pGLO digest	5 kb
N- Ndel pGLO digest	3.3kb, 1.8kb and 0.2kb
D- EcoEI and Ndel pGLO double digest	2.8 kb, 1.8kb, 0.5kb and 0.2kb