OverviewThis protocol allows for the simple enrichment of RG fractions (albeit crude) from plant samples. #####%%%%% output "RG lysis buffer" are used in line4
{"action": "allow", "output": "RG lysis buffer", "reagent": ["RG fractions (albeit crude)", "plant samples"]}
dissolved by continuous centrifugation with RG lysis buffer
{"action": "merge", "output": "", "reagent": ["RG lysis buffer"]}
The enriched RG fractions can be subsequently subjected to RNA-seq (Figure 1).imgsrc:https://en-cdn.bio-protocol.org/attached/image/e4212/-21-4212-.

Perform seed sterilization on a clean bench (wipe down with 70% ethanol before use).
{"action": "perform", "output": ""}
RNA granule enrichmentGrind 2 g of seedlings into a fine powder in liquid nitrogen using a precooled mortar .
{"action": "grind", "output": "", "device": ["a precooled mortar"], "reagent": [""], "volume": [""]}
discard the supernatant #####%%%%% output "the supernatant" are not used
{"action": "discard", "output": "the supernatant", "device": [""]}
Centrifuge at 18,000 × g for 10 min at 4°C.Resuspend the pellet in 2 ml of RG lysis buffer, .
{"action": "centrifugee", "output": "", "force": ["18,000 × g"], "time": ["10 min"], "temperature": ["4°C"]}
C.Transfer the supernatant (enriched with RGs)
{"action": "transfer", "output": "", "reagent": ["the supernatant (enriched with RGs)"]}
keep it in a freezer until use
{"action": "keep", "output": "", "temperature": [""]}
RNA analysisThe final RG fraction resulting from the protocol described above can be subjected to regular RNA extraction .
{"action": "subject", "output": ""}
subsequently tested for either targeted RNA analyses using RT-qPCR transcriptome-wide profiling with RNA-seq
{"action": "test", "output": ""}