Bio-Rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit - Manual (Page 18)

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Section 7
Vacuum Protocol
All steps are carried out at room temperature unless otherwise indicated.
Vacuum filtration steps should be carried out at –17 to –23 inHg for optimum
performance.
Important: If using the kit for the first time, please read Section 5, "Before
Using the Aurum™ Total RNA Fatty and Fibrous Tissue Kit," and Section 6,
"Vaccum Manifold Setup and Use With the Column Adaptor Plate" before
proceeding.
This procedure requires the Aurum vacuum manifold and column adaptor
plate (catalog # 732-6470), or any vacuum manifold with luer fittings. If the
necessary vacuum manifold is not available, follow the spin protocol in
Section 8.
Centrifugation steps can be performed on any commercially available
microcentrifuge that can accommodate 1.5 ml 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.
Procedure
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 of gram-positive or gram-negative bacteria
• 3.0 x 10 7 of yeast
• 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
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