Bio-rad Aurum™ Total RNA 96 Kit Manual do Utilizador descarregar pdf (Página 8)

Table 1. Yield (per well) of total RNA from various samples using

the Aurum™ Total RNA 96 kit.

Starting Material Avg. Yield (µg)*

Cultured cells (1 x 10

)

3T3 7–10

HeLa 11–17

 Bacterial(8x10

)

E. coli 5

B. cereus 5

Yeast (2 x 10

)

S. cerevisiae

9–11

Starting material amounts in parentheses are the maximum amounts recommended for use with the

Aurum™ Total RNA 96 kit.

 *Yieldfiguresarerepresentativeofaminimumoffourfullplateexperiments.

Reagents Used With the Aurum™ Total RNA 96 Kit

• The low stringency wash solution is provided as a 5x concentrate. Add

4 volumes (240 ml) 95–100% ethanol to the low stringency wash

solution concentrate before initial use.

• BeforeusingtheRNAlysissolution,add850µlofb-mercaptoethanol to the

solution,forafinalconcentrationof1%.

• The RNase-free DNase I is provided as a lyophilized powder. Reconstitute

theDNaseIbyadding250µl10mMTris,pH7.5(notsupplied)tothevial.

Pipet up and down briefly to mix. Do not vortex. Store the reconstituted

DNase I at –20°C in a nonfrost-free freezer.

• BacterialtotalRNAisolationwithoneRNAbindingplaterequirestheuseof

10 ml of TE (10 mM Tris, 1 mM EDTA, pH 7.5) for diluting the lysozyme. TE

and lysozyme are not supplied with the kit.

• Yeast total RNA isolation with one RNA binding plate requires the use of

100 ml of lyticase dilution buffer (1 M sorbitol, 0.1 M EDTA, pH 7.4,

0.1%b-mercaptoethanol), which is not supplied with the kit.

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Bio-rad Aurum™ Total RNA 96 Kit Manual do Utilizador Página 8