Bio-rad Aurum™ Total RNA 96 Kit Bedienungsanleitung PDF herunterladen (Seite 7)

Section 5

BeforeUsingthe

Aurum

™

Total RNA 96 Kit

Please read the following guidelines before proceeding with the total RNA

purification.

Starting Material Amounts

The Aurum™ Total RNA 96 kit is designed to process up to the amounts

indicated below (per well):

 • 1x10

mammalian cultured cells

• 1OD/ml*ofgram-positiveorgram-negativebacteria

 1OD/mlofbacteriaroughlycorrespondstoaculturevolumeof200–500µl

• 2OD/mlofyeast(

Saccharomyces cerevisiae

)

 2OD/mlofyeastroughlycorrespondstoaculturevolumeof300–600µl

Average yields obtained from these starting amounts are shown in Table 1.

Processing larger amounts of starting material may lead to well clogging and

reduced RNA purity.

 *Spectrophotometricdeterminationofbacterialoryeastculturedensityisa

REQUIREMENT for optimal total RNA isolation from these starting materials.

Todeterminethedensityofabacterialoryeastculture(OD

600

), combine 50

µlofculturewith950µlgrowthmedium(20-folddilution).Usethegrowth

medium as a blank and take the spectrophotometric reading at l = 600 nm.

Multiplythisfigureby20tocalculatetheOD

600

value of the undiluted bacterial

oryeastculture.DependingupontheOD

600

value, a specific volume of the

culture will be selected to provide an optimum amount of bacteria or yeast

for processing. To calculate the volume of culture required, use the following

equation:

 (OD

600

ofundilutedculture)**x(culturevolumeinml)=#OD/ml

 Forexample,2OD/mlofyeastwouldrequire800µlofanundilutedculture

withanOD

600

= 2.5.

 **1OD

600

is equivalent to approximately 8 x 10

bacterialcells/ml,or1x10

yeastcells/ml.

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