Week 7: Just Keep Swimming, Just Keep Swimming

For if you fail you must try and try again.

The lack of success in our cloning experiments is really hindering the progress of the project because we need these plasmids and sequences to make standards for qPCR.

After yet another failed cloning and transformation I decided to try once more using a sample we had successfully cloned in the past and a sample that we have had trouble cloning. We also deviated from our usual protocol in that when incubating the cells during the transformation, the samples were shaking at 200rpm (previously during incubation there was no shaking). This seemed to help because both the samples had many colonies grow on the plates and many of those colonies were white. Since cloning finally worked (Yay!) we also cloned some samples from a ciliate PCR and will be sending those samples over next week for sequencing.

This week we also tried some more Katablepharis PCR (amplification of both ITS1 and 28S regions) but neither worked well. We sent samples from the Kj3F/ITS4 PCR straight for sequencing and most of the sequences are Katablepharids which means these primers are fairly specific.

We also went to Astoria on Thursday and we got to see the equipment they put in the estuary to collect data, like Saturn one. It was cool to see all the different components on Saturn one and how they collect and transfer the data to CMOP.