Required Task CW1

Lab report (70% of total mark)

Remember the instructions you have been given in Level 1 concerning the overall structure, format and style of a lab report. The following notes give you guidance as to the expected content of each section. All tables and graphs need to be labelled correctly.

Title and Introduction:(20 marks)

• Title-Your own, reasonably short, but informative

• Background information; you should include:

• The properties of the plasmid pUC19
• Restriction digest and ligation reactions
• Transformation procedure and selection of transformants.
• Explanation of the molecular basis of blue /white colour selection to identify recombinants.
• Reference your work: make sure that you cite your sources of information correctly in the text
• Aim of the experiment

Method

• No marks are awarded for this section,however, a reference to the lab schedule/Coventry University is needed.

Results:(20 marks)

Remember that Results must have a descriptive text, as well as Figures and Tables.

1. Restriction Enzyme Digests – Presentation and Tabulation

Give a general description of the results

1. Present the photo of your restriction digests before the ligation (Labelled).
2. Using the image of your own gel (or if this is not good enough the “ideal” gel), prepare a table containing the necessary data to generate a calibration curve allowing you to estimate the size of the fragments in the restriction digests.
3. Use this data to prepare a graph on semi-log paper (provided) and attach to your write up in the appendix.
4. Prepare a table showing distance migrated and calculated sizes of fragments of each digest of B1 (Blue colony) and W1 & W2 (White colonies) attach calibration curve in appendix.
5. Show your nanodrop results with labelling.
6. Transformation
7. How many colonies were observed on each of your transformation plates?

Present a summary table of the colony count data.

1. Calculate the transformation efficiency for your experiment

Discussion:(30 marks)

Remember to use references throughout.

Restriction Enzyme Digests

1. Include all restriction fragment sizes relating to insert. Has your gel run as expected? Explain why.
2. What is the insert size according to your DNA fragment? Give your rationale

Transformation

• Why is it good laboratory practice to plate different dilutions?

1. Explain your transformation efficiency. Is it what you have expected? How could you improve your efficiency? (compare to literature).

Interpretation of your results

1. What are your findings regarding blue/white selection and antibiotics selection?
2. Discussion of the evidence that leads you to the conclusion that your experiment was successful.
3. Evaluation of your results.

Conclusion

Summary of your findings

References

Reference all work used (do not write a bibliography – reference only what you have cited)

Use the Coventry University Harvard style for your referencing

Style

Third person, past tense

Spelling / grammar

(Proof read your work)

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