BC21C Lab Report Guide 2022

Analysis of E. coli XL1 Blue Transformed Cells

Due: Sunday, March 4, 2022

Title: (1 mark)
Must be a single statement combining the titles from the two experiments done

Aim: (2 marks)
Clearly stated objectives that encapsulate both experiments.

Abstract: (10 marks)

Please recall that an abstract is a SUMMARY of the lab experiment and should
include:
• Objective
• Summary of method
• Summary of results
• Conclusion

Method: (5 marks)
• Cite the lab manual, including pages
• Changes made

Results: (10 marks)

Tabulated results for biological analysis of transformants pated on LB agar with and
without ampicillin and LB amp broth.

Discussion (30 marks)

As a guide but not limited to:
Define transformation and significance of method used
• Discuss results, what was expected to be seen on LB plate, LB broth with amp, Lb
plate with amp
• Explain the evidence used to come to that conclusion
• If the experiment was not successful, explain why
• Factors affecting transformation
• How effective is the method in determining the success of the transformation?
• Why is this method not efficient for differentiating between transformants with
pTrc99A and pTc99A-topA/CysB
• Limitations
Conclusion (2 marks)
Must answer aim using results obtained
References (5 marks)
Must have at least 3 references, APA format

Presentation and Grammar (5 marks)

Questions:
Q1, Page 136 (10 marks)

TOTAL- 80 MARKS
 BIOL2312 RESULTS 2022
EXPERIMENT 10: TRANSFORMATION

The drive link video for BIOL2312 Experiment 10: Transformation

https://drive.google.com/drive/folders/16CNyVFwV4UTG7HqlP_Lk85aCdAkXrNAG?
usp=sharing

The video is muted, but here are the steps shown:-

The XL1-Blue competent cells were already prepared. They are on ice, separated
into 100 ul aliquots.
DNA used for the transformation: Tube 2 (Post Digestion) and Tube 3 (Post Ligation)
There is also a negative i.e. No DNA, just cells.

Step 1. Addition of 10 ul of each DNA to separate tubes.

Step 2. Incubate on ice for 30 mins
Step 3. Heat Shock at 42C for 90 secs
Step 4. Add 200 ul of LB Medium to tube
Step 5. Incubate at 37C for 30 mins.
Step 6. Centrifuge for 1 min. Pour off supernatant.
Step 7. Add 100 ul of LB Medium and resuspend pellet
Step 8. Spread 100 ul on LB Amp Agar Plate. Incubate 24 hrs.

PLEASE SEE RESULTS BELOW

Negative Control
Tube 2 (Digested Plasmid & Lambda ZAP)
Tube 3 (underwent ligation protocol)
Positive Control

NB. The cell concentration in the positive control was too high, even following dilution. This resulted

in the formation of a bacterial lawn, instead of isolated colonies.

 Experiment 11: Analysis of Transformants

Here is the drive link for the accompanying video for Exp 11:
https://drive.google.com/drive/folders/16CNyVFwV4UTG7HqlP_Lk85aCdAkXrNAG?usp=sharing

Tube 3 (underwent ligation protocol)

LB Agar plate with no Ampicillin
LB Agar Plate with Ampicillin
Inoculated tubes of Broth