Here, we present a protocol for the isolation of RNA, DNA, and protein from the same sample, in an effort to reduce variation, improve reproducibility, and facilitate interpretations.
Total time
~1–2 hours per sample
Model organism
Caenorhabditis elegans
Steps
1
Collect C. elegans samples for extraction
Harvest and prepare C. elegans samples according to experimental design. Ensure proper sample collection to establish baseline material for parallel macromolecule extractions.
▶ 00:33
2
Isolate nucleotides and proteins together
Apply combined extraction protocol to isolate RNA and DNA while preserving protein from the same C. elegans sample. This unified approach minimizes sample-to-sample variation.
▶ 01:31
3
Evaluate mRNA quality via RT-qPCR
Perform reverse transcription and quantitative PCR on extracted RNA to validate nucleotide extraction efficiency and confirm mRNA integrity.
▶ 07:01
4
Assess macromolecule extraction results
Review and compare yields and quality of RNA, DNA, and protein fractions obtained from identical starting samples to confirm reproducibility.
▶ 08:12