Bio-Rad Aurum™ Total RNA 96 Kit - Manual (Page 9)

• 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 ∆A
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.
Elution Guidelines
• Apply elution solution directly to the membrane stack at the base of each
RNA binding plate.
Ribonucleases
• Although the components of this kit are provided free of contaminating
ribonucleases, great care must be taken not to contaminate the solutions or
the RNA binding plates. Gloves should always be worn when handling RNA
and should be changed frequently. Proceed through the RNA isolation as
quickly as possible with care.
• Solutions that are prepared by the user (e.g., TE) should be treated with
diethyl pyrocarbonate (DEPC) to inactivate RNases. Add 1 ml DEPC per
liter (final concentration 0.1%) of solution to be treated, mix thoroughly, and
incubate the solution at 37°C for 1 hr, or at room temperature overnight.
Autoclave the solution to remove the DEPC.
Note: DEPC is destroyed by primary amines (e.g., Tris). If a solution
containing a primary amine will be DEPC-treated, omit the amine in
preparing the solution. Perform the DEPC treatment as described above,
and add the amine to the autoclaved solution once the solution has cooled.
• Nondisposable, nonautoclavable plasticware should be rinsed with 0.1 M
NaOH, 1 mM EDTA followed by several rinses with DEPC-treated water
before use.
• Glassware and other autoclavable items may be treated using the method
described above for nonautoclavable plasticware, or by baking for 4 hr at
300°C.
• Work surfaces and micropipettors should be kept clean and wiped
periodically with an RNase removal reagent.
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