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Analysis of the project
Background Information
E.coli Transformation
Identify pBR322 in E.coli
Plasmid DNA Isolation
Calculate the copy number

본문내용

Put pBR322 into a E.coli strain and amplify it.
Try to purify the plasmid DNA to get at least 500ug of pBR322 DNA
- Analysis of the project
E.coli
E.coli
>
1
Design
rocedure
2
Confirm whether you have the correct plasmid
3
Calculate the copy number of pBR322.
Background Information
E.coli
Bacterial Transformaion
- Containing about 4000 genes
on a single circular
chromosome
Only contains single copy of
each gene
- Not harmful
large population
in a small volume
(5 x 109/ml)
Background Information
Plasmid
Bacterial Transformaion
Corcular molecule of double stranded DNA
Self replication genetic elements
Size varies from 1 to over 200kbp
Endow their hosts with useful packages of DNA
Copy Number
Copy number
The number of plasmid in a cell
Plasmids vary widely in their copy number per cell, depending on their origin of replication which determines whether they are under relaxed or stringent control. depending on the size of the plasmid
Its associated insert
- Background Information
Background Information
pBR322
One of the first vectors to be developed and the ancestor of many vectors still used today for cloning
Two marker genes that code for resistance to the antibiotics tetracycline and ampicillin
pBR322

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