ABI Sequencing and Genotyping Services
Sequencing Services
- Full Sequencing Service
- Capillary Separation Service
- Sequencing Plate Service
- Capillary Separation Service with reaction cleanup
- Plate Service with reaction cleanup
Genotyping Services
Sequencing Services
The Sequencing Service is run using a capillary ABI3730 Genetic Analyzer, from Applied Biosystems Inc.
The service offers customers a ‘Full Sequencing Service’, ‘Sequencing Capillary Separation Service’, or ‘Sequencing Plate Service’.
Full Sequencing Service
The Full Sequencing Service provides customers with the labelling of their products using cycle sequencing PCR, sequencing reaction cleanup using the X-terminator® system to remove unincorporated fluorescent ddNTPs, capillary analysis on the ABI3730 DNA Analyzer, and the analysis and uploading of sequencing data to the Genome Service ABI server for customer retrieval. There is no 96 well plate pricing for the “Full Sequencing Service”. If you are submitting 96 sample in a 96 well plate for the “Full Sequencing Service”, then you will have to pay “Full Sequencing Service” individual sample prices.
Sequencing Capillary Separation Service
The ‘Sequencing Capillary Separation Service’ covers only capillary separation on the ABI3730 DNA Analyzer, and analysis and uploading of sequencing data to the Genome Service ABI server for customer retrieval. With theCapillary Separation Service the customer is responsible for the fluorescent labelling of products, and cleanup of products to remove unincorporated fluorescent ddNTPs.
Sequencing Plate Service
The 'Sequencing Plate Service' is similar to the 'Capillary Separation Only Service' in that the customer is responsible for the fluorescent labelling of products using cycle sequencing PCR, and cleanup and precipitation of products to remove unincorporated fluorescent ddNTPs. However, the customer can do their sequencing reactions in a 96-well reaction plate, and submit the dried down plate of sequencing reactions to the Massey Genome Service for capillary separation.
The MGS is now adding a control sample onto each “Capillary Separation Service with Reaction Cleanup” and “Plate Service with Reaction Cleanup” runs. If you are planning to use the “Plate Service with Reaction Cleanup”, please only submit 95 samples and leave well H12 empty for the MGS to add a control.
Sequencing Capillary Separation Service with reaction cleanup
The ‘Sequencing Capillary Separation Service with reaction cleanup’ is similar to the ‘Sequencing Capillary Separation Service’ which covers capillary separation on the ABI3730 DNA Analyzer, and analysis and uploading of sequencing data to the Genome Service ABI server for customer retrieval. The Massey Genome Service will also cleanup your sequencing reactions using the ABI X-Terminator Cleanup method, which will remove the unincorporated fluorescent ddNTPs. You will need to set up your sequencing reactions and run them through the PCR amplification process. Please supply 10µl PCR amplified sequencing reactions in either 0.2µl tubes or 0.2µl strip tubes. For more information on the ABI X-Terminator Cleanup method, please refer to http://www.appliedbiosystems.com.au/
Sequencing Plate Service with reaction cleanup
The ‘Sequencing Plate Service with reaction cleanup’ is similar to the ‘Sequencing Capillary Separation Service with reaction cleanup’, above, but in a 96 well plate. The Massey Genome Service will cleanup your sequencing reactions using the ABI X-Terminator Cleanup method, which will remove the unincorporated fluorescent ddNTPs. You will need to set up your sequencing reactions in a 96 well plate and run them through the PCR amplification process. Please supply 10µl of your PCR amplified sequencing reactions in a 96 well plate and enter the samples into the online spreadsheet in the following order, A01, B01, C01...........H01, A02, B02, C02............H02, and so on, up to A12, B12, C12 .........H12.
Genotyping Services
The Genotyping Service is run using a capillary ABI3730 Genetic Analyzer, from Applied Biosystems Inc. The genotyping service offers fragment analysis of microsatellites.
The service offers customers either a ‘Genotyping Capillary Separation Service’ or ‘Genotyping Plate Service’ for Microsatellite Analysis.
Genotyping Capillary Separation Service
The ‘Genotyping Capillary Separation Service’ covers only capillary separation of your fluorescently labelled fragments on the ABI3730 Genetic Analyzer. With the ‘Capillary Separation Only Service’ the customer is responsible for the fluorescent labelling of fragments.
Genotyping Plate Service
The ‘Genotyping Plate Service’ is similar to the ‘Genotyping Capillary Separation Service’ in that the customer is responsible for the fluorescent labelling of fragments. However, the customer will pool their products in a 96-well reaction plate, and submit the plate of fluorescently labelled reactions to the MGS for capillary separation.
The MGS will provide customers with the 96-well reaction plates at no charge. They will be sent to you in packs of 20 plates.
Please contact the MGS by e-mail to Richard Fong (r.fong@massey.ac.nz) if you want to use our ‘Sequencing Plate Service’, and we will send you the plates. If you do not fill all 96-wells in the plate with genotyping reactions, you will still be charged the full plate price.
Please note: The Genotyping service is capable of providing SNP Analysis if there is a great enough demand for this service. Please contact the MGS if you are interested in this service by phoning 06 350 5378 or e-mail Lorraine Berry (l.berry@massey.ac.nz) Charges will be negotiated on a per customer basis.
For more information on ABI sequencing, please visit Applied BioSystems (ABI) website at: http://www.appliedbiosystems.com.au/
