Bio-rad Aurum™ Total RNA Fatty and Fibrous Tissue Kit Manual de usuario descargar pdf (Pagina 34)

Problem Possible Cause Recommended Solution

Low RNA A

260

/A

280

Some of the white Leave some of the

ratio interphase and the aqueous phase solution

(continued) organic phase behind to avoid

(containing proteins) transferring the white

were transferred with interphase with the

the aqueous phase aqueous phase (see

during aspiration into step 7 in the protocol)

a new tube

Starting material is After the sample

high in fat, proteins, disruption step,

polysaccharides, or centrifuge the lysate at

extracellular material, 12,000 x g for 10 min

causing RNA eluate at 4°C to pellet any

to be impure 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

Wash solutions and Make sure that the pipets

elution solution were that are being used for

contaminated with RNA preparation are not

proteins and other used for protein and DNA

contaminants applications

The solution used to A

260

/

280

may vary based

dilute the RNA for on the pH of the

spectrophotometric solution used to dilute

reading has a low pH RNA samples. To get

(less than pH 6.5) more accurate and

consistent A

260

/

280

values, dilute your RNA

samples with a solution

that has a pH within the

6.5–8.5 range

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