seventh week

Monday 7.4

THE LIZARDS HAVE RETURNED.

This week, Anisa and I continued to implement various miRNA isolation protocols in hopes of improving our miRNA quality to reach the 2.0-2.2 benchmark for our A260/280 and A260/230 values.

On Monday, Nikhita, Samhita, and I went to A2B for dinner, where I ordered their plain dosa.

Tuesday 7.5.2022

Today, Anisa and I each ran the kit protocol according to Dr. Gopinath’s suggestions, omitting DNAse I digestion and modifying the elution step with nuclease-free water heated to 60 degrees C. Since moving most of the lab procedures to PHRII, it has greatly reduced our need to go back and forth between JSS Medical College and PHRII, but there are still a few equipment limitations.

Wednesday 7.6.2022

Today, I accompanied Samhita for five of her interviews among the rural ASHA workers and Anganwadi workers. Since her interview consists of multiple Likert scale questions, I was able to follow along with an English translation of her questionnaire. Between interviews, we stopped for some onion pakoras!

Interesting enough, Samhita’s last few interviews were conducted at the same village that I had previously visited with Nimitha and Divya weeks ago! Since I was not able to comprehend much of their interviews since they were all conducted in Kannada and recorded on the spot, it was really helpful that Samhita’s interviews included more visual aids, which made for a more informative experience.

Thursday 7.7.2022

Today, I ran four miRNA isolations: two following a modified TRIzol protocol in duplicates while the other two included an additional kit washing step after isolation using TRIzol. This turned out to be a whole-day affair, as each isolation involves various lengthy incubation and centrifuge steps.

Later that evening, Anisa and I met with Dr. Zhou, my lab PI back in the United States, and Tasha, my lab manager in the Zhou lab, to discuss primer design and RT-qPCR SOP’s and gain some clarity on the results we’ve obtained relating to RNA yield and quality. It was really wonderful seeing them both again, and I can’t wait to return to the US to implement what I’ve learned here for our breast cancer miRNA project!

Friday 7.8.2022

Anisa and I returned to JSS today to record the Nanodrop readings for the miRNA samples isolated yesterday. I rode on the back of her two-wheeler, which was initially terrifying at first but turned out to be quite fun after a few minutes.

The rest of the day was spent validating primer sequences and researching additional miRNA isolation modifications we could make to hopefully improve our miRNA quality.

Saturday 7.9.2022

In the evening, Nikhita, Samhita, and I went to watch the Minions movie. It was only available in English in 3D! One thing I found interesting was that movies here in India always include an intermission, which allow for people to replenish snacks. The movie was definitely nostalgic, having grown up watching the Despicable Me movies.