Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Instrukcja Użytkownika Pobierz pdf (Strona 33)

Problem Possible Cause Recommended Solution

RNA is degraded Frozen tissue samples Add PureZOL directly to

(continued) were allowed to thaw or frozen samples before

sit at room temperature they thaw. Do not let

starting materials sit at

room temperature

Cells grown in either Cells grown in monolayer:

monolayer or aspirate the growth

suspension were medium and then add

washed prior to PureZOL directly to the

lysis with PureZOL plate. No trypsinization is

necessary

Cells grown in

suspension: pellet the

cells and aspirate growth

medium, then add

PureZOL directly to the

pellet

Starting tissue sample Make sure that starting

was not immediately material is immediately

frozen, or had gone processed following

through several dissection. Alternatively

freeze-thaw cycles the starting material must

before RNA be immediately frozen

purification was after dissection. Once

performed frozen, do not subject

starting material to

freeze-thaw cycles

Low RNA A

260

280

Lysate was not Make sure to incubate

ratio incubated at room the lysate after the

temperature for disruption step for

5 min after the 5 min at room

disruption step temperature to allow

(see step 3 in complete dissociation of

protocol) nucleoprotein complexes

Ethanol contamination Prior to eluting the RNA,

of the eluate make sure to perform the

purge spin step (see step

16 in spin protocol) to

remove residual ethanol

in the wash solution

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Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Instrukcja Użytkownika Strona 33

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