This was the most challenging week thus far. The sequencing data I expected returned and showed that the mutation I intended had not worked. This stemmed from a mistake in the primer design. This taught a lesson in paying extra attention to the primer design. Some measures could be used to prevent mistakes such as shown below;
Some measures that can be easily implemented involve writing the mutated codon in lowercase text and taking the primers into benchling to double check the primer against the original sequence.
Despite the mistake, New primers were ordered and having already learnt how to introduce mutations I could now go through the procedures independently and allowed me further practice and refinement of the protocols. This week was spent completing the first mutation, condensing the previous 3 weeks of work into one week. The mutation was introduced and the sample was once again sent for sequencing. Whilst the beginning of the week was discouraging, this is the nature of research. mistakes are made and protocols do not work. despite this a lot of progress was made this week and it also showed me that failure and mistakes are a part of research and are best used as learning opportunities.
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