Bio-rad Aurum™ Total RNA Mini Kit Manual de usuario descargar pdf (Pagina 23)

Section 9

Troubleshooting Guide

Problem Possible Cause Recommended Solution

Genomic DNA Incomplete DNase I Increase DNase I digest

contamination digest time

Inactive DNase I Store reconstituted

DNase I in a nonfrost-

free freezer. Avoid

freeze-thaw cycles.

Aliquot reconstituted

DNase I for single use

only

Incorrect preparation Use only the DNase

of DNase dilution dilution solution

provided in the kit to

dilute the DNase

Excessive amount of Reduce amount of

starting material starting material used

RNA degradation RNase contamination of DEPC-treat all hand-

user-made solutions made solutions.

and/or plasticware Decontaminate all work

surfaces. See Section

5 for more details

Endogenous RNases Work quickly through

the steps prior to the

addition of lysis solution

Clogging of RNA Excessive amount of Reduce amount of

binding column starting material starting material used

Poor disruption and/or Increase intensity/

homogenization duration of disruption

and/or homogenization

Switch to more intense

disruption and/or

homogenization method

Incomplete digest with Increase duration of

lysozyme or lyticase lysozyme or lyticase

digest. Use fresh

enzyme

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Bio-rad Aurum™ Total RNA Mini Kit Manual de usuario Pagina 23