Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Manual de usuario descargar pdf (Pagina 30)

Problem Possible Cause Recommended Solution

RNA binding mini Starting material is After the sample

column is clogging high in fat, proteins, disruption step,

(continued) polysaccharides, or centrifuge the lysate at

extracellular material, 12,000 x g for 10 min

causing RNA eluate at 4°C to pellet any

impurities debris present. Transfer

the supernatant into a

new 2.0 ml micro-

centrifuge tube, leaving

behind the pellet. Avoid

transferring the excess

fat that collects as a top

layer in lipid-rich

samples. Perform this

step before adding the

chloroform

Not enough vacuum Make sure vacuum

pressure was applied filtration steps are carried

for the filtrate to pass out at –17 to –23 inHg

through the columns for optimum performance.

Alternatively, transfer the

RNA binding columns to

a 2.0 ml capless tube

and centrifuge for 60 sec

>12,000 x g

Problems that may be encountered after RNA is eluted from the

column:

Low RNA yield Excessive amount of Do not exceed the

starting material maximum starting

amount limit for the kit

(see Section 5). If

clogging occurs when

using the maximum

starting amount, reduce

the amount of material

used

Inefficient elution Preheat the elution

solution to 70°C in water

bath prior to the elution

step

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Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Manual de usuario Pagina 30