Bio-Rad Aurum™ Total RNA Mini Kit - Manual (Page 9)

5
Reagents Used With the Aurum Total RNA Mini Kit
• The low stringency wash solution is provided as a 5x concentrate. Add
4 volumes (80 ml) of 95–100% ethanol to the low stringency
wash solution concentrate before initial use
• Before using the lysis solution, add 500 µl of
b-mercaptoethanol
(catalog # 161-0710) to the solution for a final concentration of 1%
• The RNase-free DNase I is provided as a lyophilized powder. Reconstitute
the DNase I by adding 250 µl 10 mM Tris, pH 7.5 (not supplied) to the vial
and pipetting up and down briefly to mix. Do not vortex. Store the
reconstituted DNase I at –20°C in a nonfrost-free freezer
• Bacterial total RNA isolations require the use of TE (10 mM Tris,
1 mM EDTA, pH 7.5), which is not supplied with the kit
• Yeast total RNA isolations require the use of lyticase dilution buffer
(1 M sorbitol, 0.1 M EDTA, pH 7.4, 0.1% b-mercaptoethanol), which is
not supplied with the kit
• Vendors of lyticase, which is used to partially degrade the cell walls of yeast
cells, may have different definitions of the enzyme’s activity. As used in this
instruction manual, 1 unit of lyticase produces a
DA
800
of 0.001/min at
pH 7.5 at 25°C, using 3 ml of yeast suspension as a substrate in a 3 ml
reaction volume
4110133B:4110133A.qxd 1/16/2009 2:35 PM Page 5