How to Operate the MagNA Pure Compact Instrument
45
B
B
For the MagNA Pure Compact RNA Isolation Kit:
The selection of an appropriate endogenous IC is of high importance when developing a
quantitative RT-PCR assay. The IC is co-amplified with the target of interest and serves as
a control for several factors: differences in initial template concentrations between differ-
ent samples, sample-to-sample variations in the PCR, presence of PCR inhibitors or the
extent of any RNA degradation. The advantage of using an endogenous IC is that both
internal control mRNA and target mRNA is extracted from cells or tissue and reverse
transcribed together. Commonly used endogenous ICs include so-called housekeeping
gene mRNAs. (Note that Roche Applied Science offers several LightCycler® Housekeep-
ing Gene Sets for the detection of human housekeeping genes, incl. G6PDH, HPRT,
ALAS, and
2-microglobulin.) The level of expression of an appropriate endogenous IC
should not vary with the experimental conditions or treatments to be compared.
Performing a Purification Run
MagNA Pure Compact Manual.book Seite 45 Dienstag, 28. Februar 2006 4:16 16
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