GE HEALTHCARE Biacore T100, Handbook

Page: 90 / 238

5 Methods
5.6 Cycle types
90
Biacore T100 Software Handbook BR-1006-48 Edition AE
The command initiates a sequence of operations in the instrument:
1 The specified volume of Deposition solution is transferred in the
autosampler to a target position in the sample and reagent rack. Target
positions are assigned as required in the
Rack Positions dialog
(Section 4.2.6). The deposition solution should be MS-compatible, and may
be used for example to neutralize the recovery conditions (which are often
acidic) or to add trypsin or another protease to the sample for peptide
digestion. The presence of deposition solution also helps to collect the small
recovered volume reliably from the autosampler needle.
2 The Sample solution is injected over the sensor surface with the Contact
time and Flow rate as specified. The Flow path is fixed so that sample
passes through all four flow cells to maximize the amount of analyte that
binds to the surface.
3 The flow system is washed with the specified Wash solution . Distilled water
or an MS-compatible buffer should be used as washing solution.
4 A small volume (approximately 2 µl) of Recovery solution , separated from
the surrounding buffer by air segments, is injected into the flow cells. The
flow is stopped for the specified
Incubation time while the recovery
solution is in contact with the sensor surface, to allow the bound analyte to
dissociate into the recovery solution.
5 The flow direction over the sensor surface is reversed and the recovery
solution containing recovered analyte is deposited in the target position
where it mixes passively with the deposition solution.
6 Steps 2-5 are repeated for the specified Number of repetitions . This
increases the yield of recovered analyte without requiring additional
commands. The same target position is used for recovered analyte from all
repetitions. Note that only one aliquot of deposition solution is used,
regardless of the number of repetitions.