Question

sheep dna was mixed with ecor1 , whuch was isolated theough gel electrophorrsis .this was then pht i to a vector and placed into e-coli. blue and white ecoli remained

10.Complete the Claim, Evidence and Reasoning on the back side of this paper. Your CER needs to focus on stages of the 511 lab: the digest, electrophoresis and transformation and what overall scientific principles you learned from this lab experience.

Answer 1

Sheep DNA was isolated and digested with EcoRI restriction enzyme and EcoRI will cleave DNA will leads to cleavage at specific site. The cleaved product will be ligated into pre digested vector with EcoRI. DNA fragment which is inserted in between beta glactosidase gene of plasmid which resulted in disruption of betag galactosidase gene.

Now ligated vector with DNA fragment is transformed into E.coli bacteria.

After incubation, the colonies appears blue and white in color. Blue represent bacterial colonies only plasmid without ligation of DNA fragment. While white colonies shows positive clone as it have vector with DNA insert.

When cloned vector enter into bacterial cells leads to no production of beta glactosidase enzyme as it's disrupted due to insertion of gene in between of beta-gal gene hence X-gal which is colorless remains colorless and colonies appear white

While in E.coli conies where only vector is transformed having intact beta galactosidase gene and hence produce beta galactosidase enzyme which convert colorlesd X-gal into blue color and hence colonies appear blue.

This overall experiment is designed to know basic steps required for ligation which involve DNA isolation, restriction enzyme digestion, ligation, transformation and screening of cells having cloned vector.

Hope it's clear.thanks