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High-Throughput Isolation of Total RNA

isolation kit from Qiagen. RNA isolations were performed according to each kits instructions. The RNeasy 96 kit does not include DNase but does make available a supplemental protocol for DNase treatment; hence, we were able to carry out an RNA isolation with as well as without DNase treatment.

The StrataPrep 96 kit, with DNase treatment, generated RNA yields similar to the RNeasy 96 kit without DNase treatment (Table 3). When DNase treatment was included with the RNeasy 96 protocol, RNA yields decreased. The StrataPrep 96 kit samples have the highest RNA concentrations, which is especially important for analyzing small samples.

* RNA was isolated from NIH/3T3 cells grown in 96-well plates (approximately 3.8 x 104 cells/well). The numbers are averages of three samples containing four pooled samples each. Yields were determined by spectrophotometric analysis.

The alternate protocol for the StrataPrep 96 kit was also tested (additional materials required).

Yield (g/well)

Concentration (g/ml)

A260/A280

StrataPrep kit

1.0 0.02

40 1.0

2.0 0.02

StrataPrep, spin only

0.9 0.07

48 2.0

2.1
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