Consumables |
The
decision as to what particular methodology
was used at each stage was based largely
on cost; this is a function of the consumables
required. For example sequencing cleanup
can be performed on either 96 or 384 well
filter plates (among other methods) but the
cost is not proportionate to the number of
wells. Consequently it is about three times
cheaper to use a 384 well plate format. |
Simplicity |
The
aim was to distil each stage of the process
down to the most simple and automation friendly
form. This facilitated standardisation and
increased robustness both in terms of hardware
operation and user interaction with the system. |
Standardisation |
It
is desirable to standardise as many processes
as possible across the range of processes
to be carried out by the system. This reduces
the number of different protocols that are
required simplifies set-up, maintenance and
user interaction with the system and increases
robustness. |
Multiplexable |
In
comparison with the capacity of the latest
analysis platforms (e.g. ABI 3730) the current
and foreseeable throughput requirements for
diagnostic genetics was considered relatively
modest. In view of this multiplexing was
kept to a minimum maximise robustness. However,
post PCR multiplexing for analysis by CSCE
was used as this was not seen to adversely
affect the system as a whole. |
Flexibility |
The
processes were designed to be as flexible as
possible, particularly with respect to batch
sizes. This enables the system to cope with
batching regimes suitable for backlogs as well
as day to day testing.
|
Robustness |
Quality
of results and Reproducibility are crucial
considerations in design and implementation
of a high throughput pipeline for mutation
detection. Re-testing for failed or poor quality
results requires time consuming re-selection
of samples as well as non-standard batches
and procedures. This can be very and disruptive
to a high throughput system. In general sequencing
due to failed primary screen is now running
at <5%.
|