Bio-rad Aurum™ Total RNA 96 Kit Uživatelský manuál stáhnout pdf (Strana 9)

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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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Bio-rad Aurum™ Total RNA 96 Kit Uživatelský manuál Strana 9