Question

- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictich enzyme Egel, and done the resulting longest fragment into Eael site of 5 KB plasmid.

Answer 1

As per the question the size of the plasmid is 5 KB (5000bp)

At the EGe1 site, a fragment of 770bp was intorduced

so the total size of recombinant plasmid would be 5000 + 770 = 5770 kb

Now as we go for the restriction digestion of the recombinat plasmid as suggested in the question

we will ge the results as

Ladder ECORA Bam Eae 1 Eco + Bam & Each 6000bP 5770be Sooobe Sodo bp coor 4000 be 3770bps 3ooobe deoobp 2000 be 2000bp 1 200 bp lotobro Tooobe +zoop 77obl Soobe cЬР

In the first case when u will digest the recombinant plasmid with EcoR1, it will cut at two places resulting in the production of two fragments. -one fragment of 2000bp and other fragment of 3770 bp

in the second case when we digest the recombinant plasmid with Bam, it will produce a single cut, hence only one fragment of 5770bp will be obtained.

in the third case when u digest the plasmid with the Eae, it will produce two cuts on the sides of the incorporated fragment and ultimately produces the frag,ment of 770 bp and 5000 bp

in the 4th case as we use all the enzymes together, they will cut at all the sites hence fragments of different lengths will be obtained.

Hope u will understand, thank u