User Manual

Section 6

Troubleshooting

Problem Possible Cause Recommended Solution

Incomplete Lysate was not mixed Once chloroform is added,

separation of properly after adding mix tubes vigorously by

phases after chloroform (See step 3 shaking for 15 seconds. Do

centrifugation in protocol) not vortex. Let the lysate

incubate for 5 minutes at

room temperature, then mix

again before centrifuging

Lysate was not Make sure that the

centrifuged at the right centrifugation is performed

temperature at 4°C following the addition

of chloroform in order to

achieve complete separation

of the phases

Incorrect amount of For every 1 ml of PureZOL

chloroform was added used, add 0.2 ml of

chloroform

Low yield Incomplete disruption Increase the duration or

and homogenization intensity of sample

of sample disrup tion.Scale up the

amount of PureZOL used

based on sample size

Over dried RNA pellet Do not use a Speed Vac to

dry the pellet. Dry pellet

briefly at room temperature

for 5 minutes. Add DEPC-

treated water to the tube

and pipet up and down

several times to resuspend

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