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45
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TM320 · Revised 4/15
10. Appendix
10.A. User-Defined Methods
The Maxwell
®
16 MDx Instrument allows the generation and use of customized user-defined methods. You can
generate a method that optimizes purification from challenging sample types or unique applications. User-defined
sample processing times for lysis, binding, drying and elution can be set. In addition, the number of cycles can be
customized for particular samples. The new run parameters can be saved on the instrument as a user method. The
e
ff
ect of any change must be determined through user testing. The potential benefits and disadvantages of
changing any of these parameters are listed below. The user will need to optimize parameters.
Potential E
ff
ects of Changing Run Parameters
Changing Lysis Time:
This is the amount of time that the plunger mixes the sample before addition of
MagneSil
®
particles to the sample.
•
Potential Benefits:
Complete lysis of the sample is necessary to recover DNA, RNA or protein. The optimal
lysis time for a specific sample type depends on the time required to adequately lyse the samples. Some
samples will not require additional lysis time after preprocessing, while other samples will benefit from an
extended lysis time.
•
Potential Disavdantages:
Extended lysis times could result in the formation of precipitates, which may
interfere with purification and downstream applications.
Changing Binding Time:
This is the amount of time that is allowed for binding the MagneSil
®
particles
containing the nucleic acid or protein.
•
Potential Benefits:
Maximal recovery requires e
ffi
cient binding of the target molecules to the magnetic
particles. Increased binding time can increase binding of the target molecule.
•
Potential Disadvantages:
Increased binding times may increase carryover of detergents, contaminants and
other components that may a
ff
ect purity or inhibit downstream applications.
Changing Drying Time:
This is the amount of time prior to elution that the particles are air-dried.
• P
otential Benefits:
Excess alcohol from the wash bu
ff
ers can interfere with downstream applications such as
gel electrophoresis, PCR and enzymatic reactions. Increasing the drying time can remove excess alcohol from
the target.
•
Potential Disadvantages:
Overdrying can reduce recovery of the target molecules from the MagneSil
®
particles.
Changing Elution Time:
This is the amount of time that the plunger is used to mix particles in the elution
bu
ff
er.
•
Potential Benefits:
E
ffi
cient recovery of the target molecules requires removal from the MagneSil
®
particles.
Extending the elution time can enhance the removal of the target molecules from the MagneSil
®
particles.
•
Potential Disadvantages:
Excess elution time can result in the release of detergents and other components
that may inhibit the downstream applications. Also, elution bu
ff
er could evaporate during long, heated
elution times.
