Roche MagNA Pure Compact Instrument, Руководство оператора

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MagNA Pure Compact Operator’s Manual - Version 1.3 66
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3. Use of Internal Controls
3.1 Function of Internal Control
If you are going to perform PCR on the purified nucleic acid and want to have an Internal
Control (IC) for that PCR, we recommend that you add the IC to the sample before you
purify it on the MagNA Pure Compact Instrument. Adding the IC at this stage allows you
not only to estimate the efficiency of the PCR, but also the efficiency of the purification.
Note: For your convenience, many of the LightCycler® reagent kits already contain such
an IC. (See the pack inserts of these reagent kits for details).
3.2 How to program the instrument for an internal control
The IC may be added to the run of the MagNA Pure Compact Instrument in either of two
ways:
As soon as one of the samples within a purification run is using IC, an IC tube has to be
inserted in
all channels that contain sample material (as indicated on the schematic on
the screen).
Note: If you program a run with an IC, but do not place any tubes in the IC positions of
the Sample Tube Rack, the instrument will pipet Lysis Buffer through the empty IC tube
postion directly onto the instrument stage. This spilled liquid may harm the operator as
well as the instrument. If this happens, you should immediately clean the instrument (as
described in chapter C) to prevent corrosion of the instrument stage. Always use gloves
when you touch the instrument or reagents, and observe the instructions on the Material
Safety Data Sheet that accompanies the reagents.
Add the internal control directly to the sample:
When you program a run with an IC in the sample, do the following: On SAMPLE
ORDERING Screen 2, choose a protocol „without IC“ in the Protocol field and
specify „none“ in the Internal Control Volume field. In the Comment field, note
that IC was added directly to the sample.
Note: Do not use this method if the IC is naked DNA ( e.g. plasmid) or RNA,
because the control may be degraded by nucleases present in the sample.
Add the IC to one of the assigned IC positions in the Sample Tube Rack. Pipet the
IC (5, 10 or 20 µl) into a separate tube. (Use only the tubes specified in chapter C,
since only they meet the instrument specifications.) Place the control tube in the
IC row (row 2) of the Sample Tube Rack.
When you program the run with separate ICs, do the following: On SAMPLE
ORDERING Screen 2, choose the protocol „with IC“ in the Protocol field and spe-
cify the correct volume of IC in the Internal Control Volume field. The instrument
will automatically include the IC tubes in the purification process.
Note: The IC will be directly mixed with the Lyis Buffer at the beginning of the run
and will thus be protected from nucleases. Therefore, choose this method if your
control is naked DNA or RNA.
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Internal Control
MagNA Pure Compact Manual.book Seite 66 Dienstag, 28. Februar 2006 4:16 16