Training activity information
Details
Extract and quantify DNA from samples according to local protocol
Type
Entrustable training activity (ETA)
Evidence requirements
Evidence the activity has been undertaken by the trainee repeatedly, consistently, and effectively over time, in a range of situations. This may include occasions where the trainee has not successfully achieved the outcome of the activity themselves. For example, because it was not appropriate to undertake the task in the circumstances or the trainees recognised their own limitations and sought help or advice to ensure the activity reached an appropriate conclusion.
Reflection at multiple timepoints on the trainee learning journey for this activity.
Considerations
- Local procedures
- Principles of DNA extraction, quantification and purity assessment
- Impact of sample source
- Safe handling of biological materials
Reflective practice guidance
The guidance below is provided to support reflection at different time points, providing you with questions to aid you to reflect for this training activity. They are provided for guidance and should not be considered as a mandatory checklist. Trainees should not be expected to provide answers to each of the guidance questions listed.
Before action
What does success look like?
- Identify what is expected of you in relation to successfully extracting and quantifying DNA from H&I samples.
- Consider how the learning outcomes apply, specifically in relation to performing the relevant laboratory investigations and practicing in accordance with quality and accreditation standards.
- Discuss with your training officer to gain clarity of what is expected of you in relation to minimum acceptable DNA concentration and purity ratios (e.g., A260/280), and accurate operation of the quantification equipment.
What is your prior experience of this activity?
- Think about what you already know about molecular techniques, nucleic acid extraction principles, and quantification methods (e.g., spectrophotometry).
- Consider possible challenges you might face during the activity, such as low sample volume limiting yield, incomplete lysis resulting in poor purity, or unexpected issues with automated extraction equipment.
- Recognise the scope of your own practice for this activity i.e. know when you will need to seek advice or help, and from whom. You will need to seek advice from your Training Officer when required, for example if the extracted DNA exhibits purity outside the acceptable range, potentially indicating protein contamination that could inhibit downstream assays.
- Acknowledge how you feel about the reliance on precise technique and equipment performance during nucleic acid preparation.
What do you anticipate you will learn from the experience?
- Consider the specific skills you want to develop, such as troubleshooting common extraction failures or accurately assessing DNA quality and concentration.
- Identify the specific insights you hope to gain into how DNA quality affects the outcome of subsequent molecular HLA typing techniques.
What additional considerations do you need to make?
- Consult actions identified following previous experiences of molecular biology preparation steps or managing complex reagents.
- Identify important information you need to consider before embarking on the activity, such as reviewing the specific local extraction protocol steps, confirming the integrity of reagents, and ensuring the quantification device is calibrated.
In action
Is anything unexpected occurring?
- Are you noticing anything surprising or different from what you anticipate whilst extracting and quantifying DNA?
- Are you encountering situations such as:
- The DNA extraction process yielding DNA of lower concentration or purity (e.g., poor A260/280 ratio) than specified by protocol?
- Unexpected issues arising with automated extraction equipment or quantification devices (e.g., technical failure, contamination)?
- Quantification revealing unexpected results, such as inhibition or readings that conflict with sample input volume?
How are you reacting to the unexpected development?
- How is this impacting your actions? For example, are you responding to the situation appropriately? Are you adapting or changing your approach by troubleshooting or attempting re-extraction?
- Consider the steps you are taking in the moment, such as:
- Immediately verifying the quantification results by re-reading the sample or performing a control extraction/quantification
- Consulting the equipment manual or technical staff if automation or quantification issues are encountered
- How are you feeling in that moment? For instance, are you finding it difficult to determine the root cause of the poor purity? Is it affecting your confidence in proceeding independently with the next step of the molecular workflow?
What is the conclusion or outcome?
- Identify how you are working within your scope of practice. For example, are you successfully obtaining DNA of sufficient quality and quantity after troubleshooting? Or are you needing support because the low yield necessitates consultation on whether to request a re-bleed from the patient?
- What are you learning as a result of the unexpected development? For example, are you mastering an efficient technique for identifying protein contamination in DNA extracts? Or gaining insight into how reagent instability can affect DNA purity?
On action
What happened?
- Begin by summarising the key steps you took when extracting and quantifying DNA.
- Consider specific events, actions, or interactions which felt important, such as how you operated the automated extractor or interpreted the spectrophotometer reading (e.g., A260/280 ratio).
- Include any ‘reflect-in-action’ moments where you had to adapt to the situation as it unfolded, for instance, immediately performing a re-read of the sample when the purity ratio was unexpectedly low or consulting the manual when the automated extractor displayed an error.
- How did you feel during this experience, e.g., did you feel precise about measurement or frustrated by equipment failure?
How has this experience contributed to your developing practice?
- Identify what learning you can take from this experience regarding DNA extraction and quantification methods. What strengths did you demonstrate, e.g., accurate documentation of quantification metrics?
- What skills and/or knowledge gaps were evident, e.g., knowledge of nucleic acid contamination indicators or troubleshooting complex extraction failures?
- Compare this experience against previous engagement with similar activities – were any previously identified actions for development achieved? Has your practice improved in adhering to local protocol and maintaining quality standards?
- Identify any challenges you experienced, such as low yield, poor purity, or needing advice on re-extraction criteria, and how you reacted to this.
What will you take from the experience moving forward?
- Identify the actions or ‘next steps’ you will now take to support the assimilation of what you have learnt, including from any feedback you have received, with regards to improving your DNA extraction technique.
- What will you do differently next time you approach DNA extraction, for instance, by proactively performing an extra blank reading on the quantification device or checking reagents more rigorously?
- Do you need to practise any aspect of the activity further, such as manual extraction protocols or key learning outcomes related to troubleshooting quantification issues?
Beyond action
Have you revisited the experiences?
- How have your subsequent experiences of extracting and quantifying DNA since completing this specific training activity led you to revisit your initial approach or decisions during that activity? For example, how a subsequent molecular assay that failed due to low DNA purity (poor A260/280 ratio) forced you to re-evaluate the rigour of your initial extraction and washing steps during your first attempt at this training activity.
- Considering what you understand about DNA yield, purity metrics, and the requirements of downstream molecular assays now, were the actions or considerations you identified after your initial reflection on this training activity sufficient?
- How have you since implemented or adapted improvements in your extraction method for difficult samples or quantification accuracy based on further learning and experiences? For example, how you proactively implemented a mandatory check of quantification equipment calibration status based on further learning.
- Has discussing different DNA extraction methods, quality control metrics, or troubleshooting extraction issues with colleagues, peers, or supervisors changed how you now view your initial experience in this training activity? For example, how professional storytelling with a senior colleague about a time when residual protein contamination inhibited a critical PCR reaction refined your understanding of the critical nature of accurate quality control metrics during nucleic acid preparation.
How have these experiences impacted upon current practice?
- How has the learning from this initial training activity, in combination with subsequent experiences of DNA extraction, contributed to your overall confidence and competence in preparing high-quality DNA samples, particularly in preparing for general observed assessments (DOPS or OCEs) involving molecular techniques? For example, how your accumulated ability in accurate quantification and protocol adherence now enables you to manage complex molecular preparations efficiently during an assessment.
- How has reflecting back on this specific training activity, combined with everything you’ve learned since, shaped your current approach to DNA extraction and quantification?
- How does this evolved understanding help you identify when DNA quality is suboptimal, or the extraction process requires expert input and when this is beyond your scope of practice? For example, how your evolved approach means you now routinely seek expert input immediately when a sample yields insufficient DNA for the required test, recognising this requires senior guidance on re-extraction or requesting a new sample.
- Looking holistically at your training journey, how has this initial DNA preparation experience, revisited with your current perspective, contributed to your development in meeting the learning outcomes related to performing relevant investigations and practicing in accordance with quality standards? For example, how this foundational experience has supported your development in transferable skills such as working with nucleic acids and understanding molecular biology principles that will be valuable in future roles or responsibilities.
Relevant learning outcomes
| # | Outcome |
|---|---|
| # 3 |
Outcome
Perform the relevant laboratory investigations for a range of patients referred to an H&I laboratory. |
| # 6 |
Outcome
Practice in accordance with quality and accreditation standards. |
| # 7 |
Outcome
Employ safe working practices to maintain laboratory equipment, prepare, handle and store laboratory reagents and patient samples. |