Listed here are the steps you would take to clone the idiosynchratase gene form

Question

Listed here are the steps you would take to clone the idiosynchratase gene form the hoot owl. Put these
into their appropriate order.
a) Ligate hoot owl genomic DNA into phage vector.
b) Expose nitrocellulose disks to UV radiation and baking.
c) Extract genomic DNA from hoot owl cells.
d) Perform autoradiography.
e) Produce a labeled probe to the idiosynchratase gene.
f) Completely digest phage vector DNA with HindIII.
g) Place nitrocellulose disks onto agar surface to transfer plaques.
h) Incubate DNA probe with nitrocellulose disks.
i) Distribute bacteria onto petri dishes containing agar and nutrients and allow growth of plaques.
j) Partially digest hoot owl genomic DNA with HindIII.
k) Infect bacteria with phage vector.

Explanation / Answer

i) Distribute bacteria onto petri dishes containing agar and nutrients and allow growth of plaques

ii) Extract genomic DNA from hoot owl cells.

iii) Partially digest hoot owl genomic DNA with HindIII.

iv) Ligate hoot owl genomic DNA into phage vector.

v) Infect bacteria with phage vector.

vi) Completely digest phage vector DNA with HindIII.

vii) Produce a labeled probe to the idiosynchratase gene.

viii) Incubate DNA probe with nitrocellulose disks.

ix) Place nitrocellulose disks onto agar surface to transfer plaques.

x) Perform autoradiography.

xi) Expose nitrocellulose disks to UV radiation and baking.

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