Sam extracted RNA using the Quick DNA/RNA Microprep Plus Kit (ZymoResearch) (post). I don’t think there is currently enough RNA in the libraries to reach the requiremets for NWGC… details in post.
Today I quickly joined his qubit result file with the hemolymph sampling data (hemo_table.csv) in R.
The resulting file: hemo_qub-for-libs.csv
The goal is to have 12 libraries accounting for each infection status and temperature treatment. Each pooled library needs to have at least 1000 ng of RNA in a sample of ~50ul. (GitHub Issue #184)
Pooled libraries to create:
| Library Number | Sample Day | Infection status | Temp trtmnt |
|---|---|---|---|
| 1 | 9 | 0 | NA |
| 2 | 9 | 1 | NA |
| 3 | 12 | 0 | Amb |
| 4 | 12 | 1 | Amb |
| 5 | 12 | 0 | Cold |
| 6 | 12 | 1 | Cold |
| 7 | 12 | 0 | Warm |
| 8 | 12 | 1 | Warm |
| 9 | 26 | 0 | Amb |
| 10 | 26 | 1 | Amb |
| 11 | 26 | 0 | Cold |
| 12 | 26 | 1 | Cold |
*Note: the temperature treatments didn’t start until after the Day 9 sampling day.
As of now, I don’t think the pooled samples would have enough RNA to meet the requirements… for example, the samples that fall under Library 1 add up to 79.9 ng/ul.
I may need to check this again when I’m not sick.