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Recombinant DNA, (Able to accept foreign DNA, 5.Small in size
-Can carry…
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- Able to accept foreign DNA
5.Small in size
-Can carry relatively large DNA inserts
-Easy to work with(convenient size for physical and manipulation
2, Has multiple cloning sites (MCS)
✓ Short segment of DNA that contains many restriction sites
✓ Has restriction sites for many restriction enzymes
✓ Allowing for easy insertion of foreign DNA
✓ Also known as polylinker site/region
- Able to replicate freely/independently in the host cell
-Have origin of replication (ori)to initate DNA synthesis
-Ori is important for the ability of plasmid to be copied (amplified)by bacteria
- Possess selectable genetic marker
-e.g. ampicillin resistance gene (ampR),IacZ gene
-To allow the transformed cells/cells containing recombinant DNA to be readily identified ( screening purpose)
- Gene cloning is a process of producing identical copies of DNA
- Both target gene and plasmid DNAs are isolated
- The target gene are amplified using PCR
- Targeted gene and plasmid DNAs are cut
- Using the same restriction enzyme
- To produce sticky ends
- Targeted gene and plasmid DNAs are purified
- Targeted gene amd plasmid DNAs are inserted using DNA ligase
- Forming recombinant DNA and transformed into (competent) DNA recombinant undergo transformation process
- Amplification process occurs
- (Blue-white) screening for white clones
- Amplification of white clones
A DNA molecule made in vitro with segments from different sources or by combining genes from different organisms.