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BluePippin Operations Manual 460013 Rev F
8-1
8
Programming Size Selections
– Tight,
Range, and Timed Modes
8.1 Tight Mode
– programming minimum size range collections
The
“Tight” programming mode is a method for selecting DNA size range collections by
entering a target base pair value in software. The system will collect the minimum fragment
distribution that is possible, centered on this base pair value.
For size reference, either internal DNA standard or external DNA markers (run in a separate
lane) may be used, depending on cassette type and/or application. Cassette kits for smaller
DNA size range collections (90-1500 bp) use internal standards and larger collections (>2
kb) require use of one cassette lane to run and external reference marker. Assignation of
reference DNA is detailed in
Sections
7.5
and
7.6
.
In the protocol editor:
1.
Select an appropriate cassette type for size selection
, if a new protocol is being
created.
2.
Assign the internal or external reference DNA
.
a. For cassettes using
internal standards
:
i. press
the “USE INTERNAL STANDARDS” button.
b. For cassettes using an
external marker
lane:
i. enter
the lane into which the marker will be loaded, into the “Reference
Lane” field (“1” is the default value)
ii.
press the “APPLY REFERENCE TO ALL LANES” button.
3.
Select
the “Tight” programming mode
. Click the “Tight” button for every lane in which
the “Tight” fragment range will be collected. If using an external marker, make sure no
collection modes are selected for that lane.
4.
Enter a value in for the “BP Target”
. This value should represent the median value for
the desired size range to be collected. The actual range to be collected will auto-fill the
“BP Start” and “BP End” fields.