FastRNA®, RED Kit
Total RNA Isolation from Yeast, Fungi, and Algae

The FastRNA RED kit uses the FastPrep machine for rapid isolation of total RNA from yeast, fungi, and algae. The lysing matrix contains medium sized ceramic particles that are agitated rapidly in the FastPrep to achieve complete lysis of yeast, fungal, and algae cells. The presence of chaotropic reagents during lysis ensures immediate stabilization of RNA. The stabilized RNA is purified by organic extractions and precipitated by isopropanol. The kit contains enough reagents for 100 total RNA preps beginning with 25-50 mg cell pellet. A preparation is completed in less than one hour.

The protocol in brief:

1. The following reagents are first prepared from solutions present in the kit:

   a. phenol acid reagent (PAR);

   b. chloroform/isoamyl alcohol (CIA);

   c. DEPC-treated isopropanol (DIPS);

   d. salt/ethanol solution (SEWS);

2. PAR, CIA and Chaotropic Stabilizing Reagents are added to a FastPrep RED tube containing lysing matrix;
3. Add up to 25-50 mg of wet cell pellet;
4. Process in FastPrep instrument for 20-40 seconds;
5. Spin in a microcentrifuge to separate phases;
6. Add additional CIA, vortex to mix, and centrifuge to separate phases;
7. Transfer upper phase to a separate tube and add DIPS to precipitate RNA; centrifuge;
8. Wash pellet by resuspension in SEWS and centrifuge
9. Based on expected use, dissolve RNA pellet in one of the three kit-supplied RNase-free solutions.

FastRNA®,  RED Kit
Cat. No.	Description	Size/Vol.
6030-600	FastRNA Kit, RED	100 Preps
Kit Contents
6030-601	FastRNA Tubes with Matrix	100 Tubes
6030-602	CRSR-Red	55 ml
6020-603	*PAR	16.5 ml
6020-604	**CIA	2.5 ml
6020-605	DIPS	1 ml
6020-606	SEWS	13 ml
6020-607	SAFE	12 ml
6020-608	SAFEE	12 ml
6020-609	SSAFE	12 ml
6020-610	BBFXE	200 µl
6020-611	LiCl	5 ml
User Supplies: *phenol, **chloroform

Yeast RNA

Total RNA isolated by the FastRNA RED Kit from 2 x 107 yeast cells in log phase growth. Lane 1: Saccharomyces cerevisiae Lane 2: Pichia pastoris