Big data from small tissues: extraction of high-quality RNA for RNA-sequencing from different oilseed Brassica seed tissues during seed development

Table 1 Amount of ground tissue used for RNA extractions and range of RNA Integrity Number (RIN) obtained for different Brassica napus and B. oleracea tissue types, n = 15

Tissue type	Amount of tissue	RIN
Ovules	6 mg	8.0–10
Ovary wall	20 mg	9.9–10
Heart stage embryo (HSE)	20–25 µl^*	8.0–9.4
Heart stage endosperm (HSEnd)	100 endosperms	7.3–9.4
Heart stage seed coat (HSSC)	2–3 mg	7.0–9.4
Torpedo stage embryo (TSE)	10–15 µl^a	7.7–9.3
Torpedo stage endosperm (TSEnd)	50 endosperms	7.0–8.4
Torpedo stage seed coat (TSSC)	2–3 mg	7.0–8.4
Green stage embryo (GSE)	50 mg	9.9–10
Green stage endosperm (GSEnd)	2–3 mg	7.8–9.5
Green stage seed coat (GSSC)	2–3 mg	8.0–9.2
Mature stage embryo (MSE)	50 mg	9.7–10
Mature stage seed coat (MSSC)	2–3 mg	7.7–9.3

^a = 100 HSE were ground in 250 µl of RB buffer from E.Z.N.A. Plant RNA kit and an aliquot of 20–25 µl was used for RNA extraction. An aliquot of 10–15 µl was used for RNA extraction of 50 TSE

ISSN: 1746-4811