The first task of the day is to check the results of the work we did with the Genomikon kit for constructing a plasmid for insertion into a bacteria. Some growth was seen on the LB plates and the LB/CHLO plates, but unfortunately there isn’t much glowing to be seen yet. Orkan kindly explained the various reasons for the lack of results at this point;
1. Bacteria have 4 phases in their growth cycle – lag, log, stationary and death. The best phase to use is the log phase in which optimal growth takes place. The fact that we were using bacteria plated 3 or 4 days previously meant we had missed this phase.
2. Due to variations in plasmid length, strain characteristics and protocol, not every bacteria strain is competent to take in the every type of plasmid. We used a strain not suggested by the kit and paid the price.
3. Maybe we didn’t add enough bacteria to the transformation buffer, this reducing the chances of success.
4. 24 hours is usually regarded as the minimum time to see meaningful results, and we observed them after only 16 hours.
A question we should maybe put to ourselves is whether, in the working environment and goals we had, knowing and practicing a protocol is more important than seeing success of that protocol. Is knowing through doing more productive than knowing by seeing?