𝐖𝐈𝐋𝐃𝐄𝐒𝐓 𝐃𝐑𝐄𝐀𝐌𝐒 | 𝐆𝐢𝐥𝐦𝐨𝐫𝐞 𝐆𝐢𝐫𝐥𝐬

54| The Lakes.

54 | The Lakes.

| Sage's POV |

≻───── ⋆✩⋆ ─────≺

"Ugh," I groaned, "I feel like my heads gonna explode."

"Everything okay?" Jess asked, pouring me a cup of coffee. "Not at all." I replied.

"Anything I can do?" He leaned against the counter, his elbows holding him up.

"Depends," I smiled, looking up. "Do you know anything about optimizing environmental DNA detection methods while also analyzing the presence of river otters in the Northeast?"

"Is Helen Keller a real person?" He countered.

"I'll take that as a no," I sighed.

"School thing?" He tilted his head. "Project. The whole class if working on it." I nodded. "I've already spent a week on it and I feel like I've done nothing."

"What have you done so far?"

"The first part of six. We had a list of species for sequencing using common mammal species near the areas of collection. We made a list of all mammals in the Northeast of the US, including species from various families. Then, we all collected DNA sequences for each of the species and analyzed for similarities. A phylogenetic tree was generated to show relation and similarities among species." I explained.

"Holy fuck," he muttered.

"Yeah, you're telling me."

≻───── ⋆✩⋆ ─────≺

Part four of this project from hell, was filtration. Definitely the easiest one so far. Two filtration protocols were used for different rivers, hand-pumped portable filtration (Protocol #1) and vacuumed in-lab filtration, using the same process but different materials.

Not too bad, but I did find myself wishing was back studying for that study language test I was so stressed about months ago.

I'd much prefer that, over this.

≻───── ⋆✩⋆ ─────≺

Part four, methods of extraction and isolation of samples proceeded for subject one, while samples for subject two were duplicated and tested twice. DNA was extracted from the filters, containing chemicals C1-C6 and PowerBead tubes. The MoBio protocol was followed for extraction and isolation.

Luckily, this one was mostly waiting. A blessing and a curse if you ask me.

≻───── ⋆✩⋆ ─────≺

Part five, two PCR amplifications in total were completed for each of the sample sets.

One: necessary to detect DNA using the broad-range primer, and two: nested PCR, making the data 100-fold more sensitive and specific.

After mixing each sample with a chemical master mix (developed using H2O and a target species primer for each of the four species), they were then placed in an Eppendorf PCR Cycler.

For the second round of amplification, the same procedure was used, but with multiple different master mixes.

≻───── ⋆✩⋆ ─────≺