Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit - Manual (Page 24)

Section 8
Spin Protocol
Important: If using the kit for the first time, please read Section 6, "Before
Using the Aurum™ Total RNA Fatty and Fibrous Tissue Kit," before
proceeding.
Centrifugation steps can be performed on any commercially available
microcentrifuge that can accommodate 1.5 and 2.0 ml microcentrifuge tubes
and can spin at >12,000 x g. It is recommended that sterile, disposable
polypropylene tubes be used throughout the procedure.
Protocol
1. Measure the amount of starting material. Note that the Aurum total
RNA fatty and fibrous tissue kit is designed to process up to the amounts
indicated below (per column):
• 1 x 10 7 cultured mammalian cells grown in suspension
• One 10 2 cm plate mammalian cultured cells grown in monolayer
• 2.4 x 10 9 gram-positive or gram-negative bacteria
• 3.0 x 10 7 yeast cells
• 100 mg of animal tissue (a 4 mm cube of most animal tissue weighs
70–85 mg)
• 100 mg of plant tissue
• 50 mg filamentous fungi
Warning: Processing larger amounts of starting material may lead to column
clogging and reduced RNA purity. It is crucial that the appropriate amount of
starting material be used. For samples that are known to be rich in RNA, it is
highly recommended that less than the maximum amount of starting material
be used so that the binding capacity of the column is not exceeded. In
addition, complete disruption and homogenization of the starting material is
critical to prevent column clogging and reduced RNA yields.
2. Disrupt and homogenize the sample. Below are recommended
procedures for disruption and homogenization.
Note: Incomplete disruption will clog the column in subsequent steps and
result in reduced yields of total RNA.
Fresh Tissue
Fresh tissue can be processed in PureZOL™ immediately after dissection.
Alternatively, freshly dissected tissue can be immediately frozen rapidly in
liquid nitrogen and processed using instructions for frozen tissue. Transfer
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