Roche MagNA Pure Compact Instrument, Руководство оператора

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MagNA Pure Compact Operator’s Manual - Version 1.3 44
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Warnings and Precautions when handling the Tip Trays:
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Check that piercing tool and reaction tips are placed correctly in the Tip Trays before
use.
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Handle Tip Trays with care to prevent tips or piercing tool from falling out of the tray.
Should this happen, discard the respective tip tray and tips. Use the Tip Tray Kit to
replace missing Tip Trays.
Note: To ensure that you have entered all essential information before starting the purifi-
cation run, the software screens will guide you through the steps for programming a
purification run. At each screen, you must enter values in all parameter fields that are
marked with a "*“ and press all confirmation buttons that are marked with a "?“ before
you can go to the next screen. All fields that are not marked with a "*“ are for documenta-
tion only; you do not have to fill these in. Nevertheless, we recommend that you enter
values in these fields, since this information may later help with troubleshooting.
Controls
Always run appropriate controls with the samples, especially if you want to perform
quantification analyses of the eluted DNA samples (e.g., by LightCycler® 2.0 System PCR
assays). In order to control the complete process starting from sample preparation to
quantification analysis, perform the following controls:
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Positive Control , by using a sample material positive for your target.
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Negative Control , by using a sample material negative for your target.
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Internal Control , by adding a defined amount of a control template (e.g., plasmid
DNA) to all samples to be purified or by analyzing an endogenous nucleic acid
sequence present in all your samples.
For the MagNA Pure Compact Nucleic Acid Isolation Kit and MagNA Pure
Compact Nucleic Acid Isolation Kit - Large Volume:
The Internal Control (IC) is added prior to the purification step, then co-purified, and
amplified with your target of interest from the specimen in the same PCR reaction. The
IC concept is especially useful for enzyme-based amplification processes such as PCR,
because inhibitors present in the purified sample material might reduce efficiency of the
PCR process. In addition, the Internal Control is used to compensate for possible losses
of your target during purification.
For LightCycler® 2.0 System quantification assays use a synthetic double-stranded DNA
molecule with primer-binding sites identical to those of your target sequence, but having
a unique probe-binding region that differentiates the IC from the target-specific ampli-
con. Discriminate the signals derived from your target and the IC by performing a dual-
color HybProbe assay. For detailed information regarding the IC concept in combination
with the LightCycler® 2.0 System, read LightCycler® Technical Note 12/2000 “Absolute
Quantification with External Standards and an Internal Control” available at http://
www.lightcycler-online.com.
Performing a Purification Run
MagNA Pure Compact Manual.book Seite 44 Dienstag, 28. Februar 2006 4:16 16