Genotyping pipeline
The KBioscience genotyping pipeline is extremely robust and capable of scaling to over 100 million genotypes per year by increasing the multiples of equipment.
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Installation
The installation is carried out by experienced KBioscience staff with full on site and remote training if required, Basic pipeline will be capable of ~50,000 genotypes per day. KBioscience pipeline is a full turn key solution and includes:
- Preparation of DNA plates (daughter) for genotyping.
- Tip Regeneration
- Mastermix dispensing
- Plate Sealing
- Thermal Cycling
- Plate Reading
- LIMS
- Ancillary Equipment
- Consumables
- Warranty and technical support
Preparation of DNA plates (daughter) for genotyping.
Standard 384 plates dispenser, modified to take the KBioscience 384 and 1536 well plate formats. These are two separate stand-alone pipetting robots. Taking this approach provides redundancy and flexibility. It should be noted that the cost of the consumable tips for these robots will be inhibitory unless a tip regeneration system is utilised. At Kbiosciences we have been regenerating tips for a period of 5 years. It has proven to be highly cost effective and reliable.
Tip Regeneration
A semi-automated tip washing system will be supplied, along with tried and tested protocols for the process of tip regeneration. The system will require a deionised water supply and a compressed air supply.
Mastermix dispensing
A KBioscience modified Equator 8 channel dispenser from Deerac fluidics would be supplied. The modifications would include:
- Peltier cooled plate nest to avoid evaporation issues
- Extended tubing array fitted to allow for large volume aspiration (multiple plate dispenses)
- New 8 channel aspirate vessel fitted to allow for continuous 8 plate per aspirate dispensing
- Modified PIC central processor to upgrade the motion algorithms along with allowing KBioscience LIMS dispense lists, enabling partial plate dispensing.
- KBioscience K-Equator software for at ‘dispense’ assay and machine logging.
- Fast wash addition to increase throughput.
The modifications allow for approx 32 x 384 plates per hour dispensing or 24 x 1536 plates per hour dispensing.
Plate Sealing
A Fusion laser plate sealer will be supplied for sealing the plates. Its capacity is approximately 75 plates per hour.
The fusion requires 5 bar dry compressed air.
Thermal Cycling
Included in the installation is the supply of two Hydrocycler 16 water bath thermal cyclers. Two lower capacity instruments are selected instead of one 32 plate machine due to the flexibility, redundancy and throughput. One of the two HC16 can be started off during a production day when 16 plates are ready from the Equator, this means the time to first run is only 30 mins, as opposed to 1 hour if a HC 32 is used. The HC-16 also has the advantage of requiring no facilities modifications as it can be run from a 220v single phase supply, this is not possible from a HC32.
Plate Reading
The raw data acquisition will be carried out by a standard plate reader fed by a modified Zymark twister robot, with barcode reading enabled.
LIMS
KBioscience will install its industrial mySQL powered LIMS tracking and data analysis system.The scope of this software is too large to be covered here. The software enables all the instrumentation to be controlled from a central point with all plates, oligos, assays and samples to be tracked by 1D and 2D barcodes.
A networked server PC running Windows server 2003 will be required.
Ancillary Equipment
KBiosciences will also supply a combination of 1D barcode readers (2), 2D barcode rack scanners (1) and a 2D barcode tube reader (1) to support all tracking functions in the laboratory.
Consumables
It should be noted that the pipeline does not require the use exclusively of KBioscience consumables.
KASP chemistry
KBioscience recommend the use of its industry proven KASPar genotyping chemistry.