Molecular Biology Techniques

Question 1

What are the seven steps in the cloning of specific DNA sequence?

Answer 1

Choose host organism and vector

prepare vector DNA

cut DNA to be cloned with restriction enzymes

create recombinant DNA with DNA ligase

introduce recombinant DNA into host organism(transformation)

select host containing vector DNA

screen for clones with desired inserts

Question 2

Required features of an expression vector

Answer 2

promoter and start codon

polyadenylation signal

inducer

fused tag for identification and purification

two proteins expressed simultaneously (IREB site)

method of cloning (restriction enzymes or PCR product)

antibiotic selection