Preparation of RNA samples
Note: The quality of the RNA is essential to the overall success of the GeneChip analysis. Since the most appropriate protocol for the isolation of RNA can be source material-dependent, we recommend using a protocol that has been established for the tissues and cells being used. In the absence of an established protocol, we suggest using one of the commercially available kits designed for RNA isolation.
Isolation of RNA from mammalian cells or tissues
When using a commercial kit, follow the
manufacturer's instructions for RNA isolation. Good quality RNA has been
obtained using the kits listed below:
When using a commercial kit, follow the manufacturer's instructions for RNA isolation. Good quality RNA has been obtained using the kits listed below:
Total RNA preparation from cells: QIAGEN's RNeasy Total RNA Isolation kit.
Total RNA preparation from tissues: Invitrogen Life Technologies' TRIzol reagent.
Note: When total RNA is isolated using TRIzol, a second RNA cleanup step using QIAGEN RNeasy Total RNA isolation kit can be used. This gives much better yield of labeled cRNA.
In general, 0.1-0.5ug total RNA (high quality)at 0.1-0.4 ug/ul concentration is needed to obtain sufficient quantity of labeled cRNA for hybridization to GeneChip arrays.