Tools And Techhniques Flashcards
What do derivation enzymes do
Cut double stranded DNA ( @ recognition site). From interior
What are they used for
Cut join replicate DNA pieces to make recombinant DNA.( basis of gmo)
What is a recognition site
4-8 base pairs
Where do restriction enzymes come
From bacteria, uses them as primitive immune system
How were enzymes in bacteria used as immune system
They cut up viral DNA so they don’t replicate
What are blunt and sticky ends
Products of festivities enzymes ( blunt ends - flat, sticky ends - overhangs)
How do we join sticky ends or blunt ends together
Same restriction enzymes is needed to cut two different pieces of DNA ( for both)
DNA ligand used only ( sticky ends - h bonding )
T4 dna ligase ( bluntness )
How does DNA ligase work
Condensation reaction ( takes t water, changing the phosphodiester bond)» backbone of DNA
What are plasmids
Small circular pieces of DNA ( exist in bacteria )
Why do bacteria have plasmid
Carry genes that help convey resistance
What are plasmids used for
Make rec DNA
Vectors ( splice foreign gene into it and the use it for ur own use)
What is and an example of plasmid and rec DNA
Insulin Isolated and inserted into plasmid and then inserted in to bacteria. Bactria make more with protein synthesis
What are plasmid requirments
Origin of replication
Selectable marker ( resistance or lac Z»_space; restriction site)
Restriction enonuclease site
Explain the requirments
Copied independent then bacteria
Protected
Inserts gene to be cloned
What is selective plating
Check if transformation was successful .
amp thing at lab
