Counting cytoplasmic incompatibility factor mRNA using digital droplet PCR

Wolbachia bacteria inhabit over half of all insect species and often spread through host populations via efficient maternal transmission and cytoplasmic incompatibility (CI), killing aposymbiotic embryos when fertilized by symbiotic males. Wolbachia's cifB gene triggers CI in males, while cifA, expressed in females, rescues embryos from CI-induced lethality. In some systems, cifA also contributes to CI induction. CI strength—the percentage of embryos that die from CI—is a key determinant of Wolbachia's prevalence in host populations, and cifB mRNA levels in testes generally correlate with CI strength. Yet, cifB's rarity can hamper precise quantification, necessitating tissue pooling for reverse transcription quantitative PCR (RT-qPCR) to achieve reliable measurements, obscuring variation at the level of individual insect tissues. Here, we present four RT digital droplet PCR (RT-ddPCR) assays to count rare cifA and cifB mRNA from wMel Wolbachia in Drosophila melanogaster. These assays co..., , # Data from: Counting cytoplasmic incompatibility factor mRNA using digital droplet PCR

Dataset DOI: [10.5061/dryad.0k6djhbd8](10.5061/dryad.0k6djhbd8)

## Description of the data and file structure

All data associated with Van Vlaenderen L, Conner WR, Shropshire JD. (2025) Counting cytoplasmic incompatibility factor mRNA using digital droplet PCR.

### Files and variables

#### File: Data-S1.csv

**Description:** RNA extraction and purification measurements.

##### Variables

* Sample ID: A unique sample identifier
* Testes: Number of testes in the sample
* pre-Qubit concentration (ng/L): RNA concentration measured through Qubit before reprecipitation
* pre-Nanodrop concentration (ng/L): RNA concentration measured through Nanodrop before reprecipitation
* pre-A260/A280: Nanodrop quality ratio before reprecipitation
* pre-A260/A230: Nanodrop quality ratio before reprecipitation
* pre-A260: Nanodrop measurement before reprecipitation
* pre-A280: Nanodrop measurement before reprecipitat...,