Steven asked that I help isolate RNA (GitHub Issue) for Noah’s summer project involving heat and mechanical stress (tumbling) of adult Crassostrea gigas (Pacific oyster). Here’s a brief overview of the process:
- Frozen ctenidia tissue was kept on ice.
- Piece of tissue was placed in 500uL RNAzol RT in a 1.7mL tube and immediately homogenized using a disposable pestle.
- An additional 500uL of RNAzol RT was added to the tube, vortexed, and incubated at room temp for ~5mins.
- Insoluble material was pelleted at 21,000g for 1minute.
- Supernatant was transferred to 2mL tube containing 1mL 100% ethanol and vortexed.
Used 700uL of each sample for use in the Direct-zol mini RNA Prep Kit (ZymoResearch). Manufacturer’s protocol was followed, including DNase step.
Samples were stored in Noah’s -80C project box. RNA will be quantified tomorrow.
| Sample |
|---|
| NTC1_ctenidia |
| NTC2_ctenidia |
| NTH1_ctenidia |
| NTH2_ctenidia |
| NTHM1_ctenidia |
| NTHM2_ctenidia |
| NTM1_ctenidia |
| NTM2_ctendia |
| TC1_ctenidia |
| TC2_ctendia |
| TH1_ctenidia |
| TH2_ctenidia |
| THM1_ctenidia |
| THM2_ctenidia |
| TM1_ctendia |
| TM2_ctenidia |