Try these question~~!!
Q : What are the general procedure involve in gene cloning?
A : Gene cloning involve 5 main procdure, that are extraction of DNA, generation of DNA fragment, insertion of DNA fragment into vector, transform vector into host and screening for succesful recombinant.
Q : What is the best vector to be used in gene cloning?
A : Depending on the size of DNA fragment to be inserted. For example, inserting a large DNA fragment into plasmid may not be applicable since plasmid can only accommodate up to 10kb of DNA.
On the other hand, vector to be used also depending on types of host to be transferred. For example, Ti plasmid can be used to transfer gene into plant cell but plasmid cannot.
Q : What type of ligase is use for blunt end ligation?
A : T4 DNA ligase can be used in blunt end ligation. However, E.coli DNA ligase can only use for cohesive end ligation.
Q : What is the phenotype of successful recombinant in blue-white screening?
A : White colony. This is because the foreign DNA that inserted disrupt the gene sequence of Lac Z gene, which causing that gene malfunction and not able to produce β-galactosidase to degrade X-gal. Therefore, no blue colour dye is formed and the colony is in white colour.
A : Gene cloning involve 5 main procdure, that are extraction of DNA, generation of DNA fragment, insertion of DNA fragment into vector, transform vector into host and screening for succesful recombinant.
Q : What is the best vector to be used in gene cloning?
A : Depending on the size of DNA fragment to be inserted. For example, inserting a large DNA fragment into plasmid may not be applicable since plasmid can only accommodate up to 10kb of DNA.
On the other hand, vector to be used also depending on types of host to be transferred. For example, Ti plasmid can be used to transfer gene into plant cell but plasmid cannot.
Q : What type of ligase is use for blunt end ligation?
A : T4 DNA ligase can be used in blunt end ligation. However, E.coli DNA ligase can only use for cohesive end ligation.
Q : What is the phenotype of successful recombinant in blue-white screening?
A : White colony. This is because the foreign DNA that inserted disrupt the gene sequence of Lac Z gene, which causing that gene malfunction and not able to produce β-galactosidase to degrade X-gal. Therefore, no blue colour dye is formed and the colony is in white colour.