Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Manual de usuario descargar pdf (Pagina 32)

Problem Possible Cause Recommended Solution

Genomic DNA On-column DNase I Perform the on-column

contamination digest was not DNase I digest (see step

performed 14 in protocol)

Incomplete DNase I Increase the digest time

digest for starting materials that

are known to contain a

high level of genomic

DNA

DNase I is inactive Reconstitute the

lyophilized DNase I with

10 mM Tris, pH 7.5.

Aliquot and store the

reconstituted DNase I

enzyme in a nonfrost-free

freezer. Avoid freeze-

thaw cycles by aliquoting

the enzyme for single use

only

Some of the white Leave some of the

interphase (after aqueous phase solution

phase separation) behind to avoid

was transferred with transferring the white

the aqueous phase interphase with the

aqueous phase (see

step 7 in the protocol)

RNA is degraded RNase contamination Make sure to use

of solutions provided RNase-free plasticware.

in the kit See Section 5,

Maintaining an

RNase-free Environment,

for detailed instructions

RNase contamination Treat all user-made

of solutions supplied solutions with DEPC

by the user before use (see Section 6

for instructions)

RNase contamination See Section 6,

of plasticware and Maintaining an RNase-free

work station Environment, for detailed

instructions

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Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Manual de usuario Pagina 32