In the process of doing interlab-2023-exp1, we have met two main problems. The first one, which is also the major one, is transforming DNA successfully into E. coli cells and we have gained some our own experience, hoping to help improve exist troubleshooting protocol. The other one is the setting for measurement, which can be found only when we do the measurement. We hope we can offer some experience and help future teams.
Our tough journey is as follows and our solutions are marked below:
First, we have used DNAs in other wells (not the
experiment wells) to test the transformation efficiency
and the transformation didn’t work well. We have asked other igem teams but they didn’t do the interlab,
so we inquired the committee and Vinoo Selvarajah, the vice president of interlab, has given some useful
advice.
After that, we checked the Transformation Troubleshooting and made sure the quality of the
protocol meets the standard.
Then we have raised the concentration by transforming
half of the DNA in the well according to the
troubleshooting handbook, but it still didn’t work. What’s more, due to the lack of the time to
deadline, we have used 4 experiment wells directly this time, so we are very frustrated to see this
result.
Regarding this as the last chance, we transformed the other 2 experiment wells’ whole DNA.
To our extremely surprising, the transformation has
succeeded this time, so we transformed the last 2
experiment wells. However, one of the 2 wells has transformed successfully but the other one has
failed. To find out if the “using the whole DNA” protocol has worked and to fulfill the measurement of
all 8 experiment wells, our team member has borrowed other team’s distribution kit.
This time, we transformed all of 8 experiment wells,
but left one sample to the freezer after recovery
because we only found 7 culture dishes. We desperately found that all of 7 transformations have
failed. Then, we cultured the left one in the freezer after 2 days not seeing the 7 dishes’ results.
Strangely, it has worked, which suggests that whether it was the problem of our team member’s mistake,
or storage in the freezer after recovery step may help stimulate the antibiotic ability of the DNA.
Storage in the freezer after recovery step may help stimulate the antibiotic ability of the DNA.
It is worth mentioning that, after that, we also tried
to lower the concentration of the
Chloramphenicol to see if the transformation of half of the DNA in the well can work (our left 4
experiment wells’ DNA).
The result suggested that Chloramphenicol concentration shouldn’t be lowered and half of the DNA in
the well is not enough to be transformed.
To sum up, we think that all of the DNA in the wells
should be utilized if the transformation didn’t
work and there are also chances that the transformation didn’t work if our team member didn’t make
mistakes (which can be testified by transforming other wells).
