Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit User Manual
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Reagents Used With the Aurum Total RNA Fatty and Fibrous
Tissue Kit
PureZOL™ RNA Isolation Reagent for Sample Lysis
Use 1 ml of PureZOL for up to:
•
100 mg of tissue
•
1 x 10
7
cultured cells grown in suspension
•
One 10 cm
2
plate of cultured cells grown in monolayer
•
2.4 x 10
9
of gram-positive or gram-negative bacteria (equivalent to
3 OD•ml of bacteria)
•
3 x 10
7
of yeast (equivalent to 3 OD•ml of yeast)
Low Stringency Wash Solution
•
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
DNase I
•
10 mM Tris, pH 7.5 prepared in DEPC-treated water (not supplied) is
required to reconstitute the RNase-free DNase I that is provided as a
lyophilized powder
•
Reconstitute the lyophilized DNase I by adding 250 µl of 10 mM Tris, pH
7.5 to the vial and mix by briefly pipetting up and down. Do not vortex
•
Aliquot and store the reconstituted DNase I at –20°C in a nonfrost-free
freezer. Avoid freeze-thaw cycles
Note: 5 µl of the DNase I stock is needed per column or prep. When the
DNase is ready to be used, it must be mixed with 75 µl of the DNase
dilution solution (provided in the kit) per column. Once diluted with DNase
dilution solution, use the DNase immediately and do not refreeze for later
use
Elution Guidelines
•
Apply elution solution directly to the membrane stack at the base of each
RNA binding mini column
Preparation of DEPC-Treated Water
•
Autoclaving of laboratory solutions and buffers used for RNA preparation
does not guarantee the complete inactivation of RNases, which can
maintain residual activity causing RNA samples to be degraded. For this
reason, solutions and buffers should be treated with diethyl pyrocarbonate
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