Roche MagNA Pure Compact Instrument - Performing a Purification Run

7
Important Information regarding the MagNA Pure Compact Instrument Operator’s Manual
2. Performing a Purifi cation Run, page 45
Current Version Changes
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-amplifi ed with
the target of interest and serves as a
control for several factors: differences
in initial template concentrations
between different 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
®
Housekeeping 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.
The selection of an appropriate
endogenous IC is of high importance
when developing a quantitative RT-
PCR assay. The IC is co-amplifi ed with
the target of interest and serves as a
control for several factors: differences
in initial template concentrations
between different 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. The level of expression of an
appropriate endogenous IC should not
vary with the experimental conditions
or treatments to be compared.
Chapter B
22051208 MagNA Pure Compact SW Update 1.1.2 Addendum 2.indd 722051208 MagNA Pure Compact SW Update 1.1.2 Addendum 2.indd 7 24.03.2009 16:05:1824.03.2009 16:05:18