Pglo Transformation Lab Report
Title of the lab: Transformation : Bacterial Genetics
Purpose of the lab: The pupose of the lab was to transfor a bacterial E. Coli by using the green flurescent protein from the jellyfish. Another important that was fferdone by making the cell competency, meaning that it will be able to take on additional DNA. This was done when the plasma was added.
Materials:
1. 37 o C water bath
2. Ice
3. Sterile transfer pipette
4. Foam tube rack
5. Transformation solution (CaCl2)
6. pGLO plasmid
7. Sterile Inoculating loop
8. 2 - LB+amp plate
9. LB+amp+ara plate
10. LB plate
11. LB broth
12. E.coli starter plate culture
Procedure:
The test tubes were labael +pGLO and another –pGLO, and placed them on a test tube.
Transfer 250uL of transformation solution CaCl2
The tubes were place in an ice bath
Transfer a colony of bacteria by using a sterile loop and mak e sure all is mixed well
Return the tube on the ice bath and redo step 4 with the –pGlo tube.
Put a loop full of plasmid DNa into the +pGLO tube and make sure you mix it well
Return the tune over the ice bath for additional 10mins.
Place tube into water bath for 50 seconds, transfer them back and incubate for teo minuted
Add 250 uL of LB nutrient agar and incubate at room temperature for 10 minutes
Transfer …show more content…
For the tube that did not show growth meanth that the E-coli was never transformed and perheps never had a plasmid introduce to it. If bacteria did grow, it meant that the E. Coli was transformed and was now ampicillin resistant. E- Coli is naturally not resistant to ampicillin, but in this case it contain an ampilicillin resistant gene, that when introduced to E. Coli allows it to grow in the presence of ampicillin. ---pGLO LB, +pGLO LB LB+Amp -pGLO LB LB+Amp+Ara showed growth meaning it had the plasmid, but the -pGLO LB+Amp did not show growth meaning that it did not have
Purpose of the lab: The pupose of the lab was to transfor a bacterial E. Coli by using the green flurescent protein from the jellyfish. Another important that was fferdone by making the cell competency, meaning that it will be able to take on additional DNA. This was done when the plasma was added.
Materials:
1. 37 o C water bath
2. Ice
3. Sterile transfer pipette
4. Foam tube rack
5. Transformation solution (CaCl2)
6. pGLO plasmid
7. Sterile Inoculating loop
8. 2 - LB+amp plate
9. LB+amp+ara plate
10. LB plate
11. LB broth
12. E.coli starter plate culture
Procedure:
The test tubes were labael +pGLO and another –pGLO, and placed them on a test tube.
Transfer 250uL of transformation solution CaCl2
The tubes were place in an ice bath
Transfer a colony of bacteria by using a sterile loop and mak e sure all is mixed well
Return the tube on the ice bath and redo step 4 with the –pGlo tube.
Put a loop full of plasmid DNa into the +pGLO tube and make sure you mix it well
Return the tune over the ice bath for additional 10mins.
Place tube into water bath for 50 seconds, transfer them back and incubate for teo minuted
Add 250 uL of LB nutrient agar and incubate at room temperature for 10 minutes
Transfer …show more content…
For the tube that did not show growth meanth that the E-coli was never transformed and perheps never had a plasmid introduce to it. If bacteria did grow, it meant that the E. Coli was transformed and was now ampicillin resistant. E- Coli is naturally not resistant to ampicillin, but in this case it contain an ampilicillin resistant gene, that when introduced to E. Coli allows it to grow in the presence of ampicillin. ---pGLO LB, +pGLO LB LB+Amp -pGLO LB LB+Amp+Ara showed growth meaning it had the plasmid, but the -pGLO LB+Amp did not show growth meaning that it did not have