-Match the observation with the best-fitting explanation in the context of our r
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Question
-Match the observation with the best-fitting explanation in the context of our recDNA experiment:
1.Cells after transformation procedure are spread-plated onto nutrient agar that includes an appropriate antibiotic.
2.X-gal and IPTG
3.Blue-white colour screening
4.Blue colonies growing on nutrient agar with ampicillin that includes X-gal plus IPTG
5.White colonies growing on nutrient agar with ampicillin that includes X-gal plus IPTG
6.Colonies growing on nutrient agar that includes ampicillin
7.Colonies growing on nutrient agar that includes kanamycin
8.Colonies growing on blood agar that show a translucent patch in the agar around them (i.e. haemolysis)
9.Colonies growing on blood agar that show no change in the agar around them (i.e. no haemolysis)
10.Colonies that grow on medium containing ampicillin, and are white on medium containing both X-gal and IPTG, and are shown to be haemolytic
choise below:
-These cells contain a plasmid that does not have the hlyA gene.
- Cells that contain original pBluescript now look different to cells that contain pBluescript that now has an insert (a piece of DNA ligated into the plasmid). - The vast majority of cells will die because only ~1/1000 cells takes up plasmid DNA
- Cells in these colonies should contain the plasmid that achieves the aim of our Prac A subcloning experiment.
- When these chemicals are in the agar then cells with a functional lacZ gene (encoding beta-galactosidase enzyme) turn blue.
- These cells contain a plasmid that is based on pK184 (could be unchanged pK184 or pK184 that now has an insert).
- These cells contain a plasmid that now has the hlyA gene.
- These cells contain a plasmid that is based on pBluescript (could be unchanged pBluescript or pBluescript that now has an insert).
- These cells contain original pBluescript (no insert ligated into the plasmid).
- These cells contain pBluescript that now has an insert (a piece of DNA ligated into the plasmid).
Explanation / Answer
1. Cells after transformation procedure are spread-plated onto nutrient agar that includes an appropriate antibiotic. - Cells that contain original pBluescript now look different to cells that contain pBluescript that now has an insert (a piece of DNA ligated into the plasmid).
2. 2.X-gal and IPTG - When these chemicals are in the agar then cells with a functional lacZ gene (encoding beta-galactosidase enzyme) turn blue.
3. 3.Blue-white colour screening - These cells contain a plasmid that is based on pBluescript (could be unchanged pBluescript or pBluescript that now has an insert).
4. 4.Blue colonies growing on nutrient agar with ampicillin that includes X-gal plus IPTG - These cells contain original pBluescript (no insert ligated into the plasmid).
5. 5.White colonies growing on nutrient agar with ampicillin that includes X-gal plus IPTG - These cells contain pBluescript that now has an insert (a piece of DNA ligated into the plasmid).
6. Colonies growing on nutrient agar that includes ampicillin - The vast majority of cells will die because only ~1/1000 cells takes up plasmid DNA
7. Colonies growing on nutrient agar that includes kanamycin - These cells contain a plasmid that is based on pK184 (could be unchanged pK184 or pK184 that now has an insert).
8. 8.Colonies growing on blood agar that show a translucent patch in the agar around them (i.e. haemolysis) - These cells contain a plasmid that now has the hlyA gene.
9.Colonies growing on blood agar that show no change in the agar around them (i.e. no haemolysis) - These cells contain a plasmid that does not have the hlyA gene.
10.Colonies that grow on medium containing ampicillin, and are white on medium containing both X-gal and IPTG, and are shown to be haemolytic - Cells in these colonies should contain the plasmid that achieves the aim of our Prac A subcloning experiment.
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