Introduction (15%)
Include some general background to microcephaly and centrosomes
Explain why a single nucleotide change caused the early termination of CDK5RAP2
State a hypothesis and the rationale of performing experiments
Aim (5%)
What is the main aim to produce in these experiments
How this fits with strategy of uses of GFP
Method/Strategy (including diagrams) 25%
Include two sub-sections:
Plan of how to detect whether CDK5RAP2E1516Ter localises to centrosomes by fluorescent microscopy
Plan of how to detect whether CDK5RAP2E1516Ter interacts with PCNT by GFP-pulldown followed by western blot
Plan of plasmid constructs that will be used for experiments
Plan of how you perform transfection of plasmids to cells
Plan of how you prepare microscope samples and image them
Plan of how you perform Western blot
Describe antibodies used for your experiments
Note that methods should be written in a way that readers can repeat the same experiments. However, for this report, no need to go into minute details of experiment i.e. 5ul of x into Y, name of equipment (Zeiss LSM880 confocal microscope)
Trouble shooting 5%
List elements that will need testing or trouble shooting in strategy and why
Controls 5%
List the controls required (positive and negative) and the rationale for their use
Alternative strategy 5%
If this strategy fails what other would you employ and why
Results 25%
Include two sub-headings for the two experiments that you did. The sub-heading should highlight summary of results, e.g. “CDK5RAP2E1516Ter does not interact with PCNT”
Present example results in terms of fluorescent microscopy image and Western blot (2 figures)
These could be “designed” or taken from research paper
“Designed” figures would take the form of expected fluorescent microscopy image or gel/blot appearance, and could be made in powerpoint or similar
Remember scale bars, nuclear staining, ladders etc
Provide figure legends
Text should describe results presented in figures
Discussion 10%
Did you achieve aims?
Put results in wider context of how the E1516Ter mutation impacts centrosome functions and disease
5% will be awarded for the quality of the presentation and accurate referencing.