The Full Sequencing Service provides customers with the labelling of their products using cycle sequencing PCR, sequencing reaction cleanup using the CleanSeq magnetic bead system to remove unincorporated fluorescent ddNTPs, capillary analysis on the ABI3730 DNA Analyzer, and the analysis and uploading of sequencing data to the AWCGS server for customer retrieval.

Samples must be supplied in either 0.2ml individual PCR tubes or 0.2ml strip tubes. If you are supplying more than 30 samples in one request can you please supply the samples in the 0.2ml strip tubes.

It is the customer's responsibility to prepare, purify, and quantify their DNA products, before sending to the AWCGS for sequencing. Information on the best methods for purifying and quantifying DNA products for sequencing refer to the section on 'DNA Template Purification and Quantification'. The AWCGS also requires customer to premix their template and primer before sending to the service for sequencing. Please refer to ‘DNA Template and Primer Concentration Requirements’ for details.

The Capillary Separation Service covers only capillary separation on the ABI3730 DNA Analyzer, and analysis and uploading of sequencing data to the AWCGS server for customer retrieval. With the Capillary Separation Service the customer is responsible for the fluorescent labelling of products, and cleanup of products to remove unincorporated fluorescent ddNTPs.

Samples must be supplied in either 0.2ml individual PCR tubes or 0.2ml strip tubes. If you are supplying more than 30 samples in one request can you please supply the samples in the 0.2ml strip tubes.

Please follow the guidelines in the sequencing kit protocols if you are doing your own sequencing reactions for the Capillary Separation Service, to get optimal results.

The sequencing kit protocols, which the AWCGS support, can be downloaded as PDF files from the section called 'Chemistry Guide Downloads'. Also refer to the section called 'Tips on Performing Sequencing Reactions'.

For cleanup of your florescent labelled sequencing products you can either follow the cleanup method in the Chemistry Guide above, which is ethanol precipitation, or use a magnetic bead cleanup method. The AWCGS Full Sequencing Service uses the Agencourt CleanSeq magnetic bead sequencing cleanup method to remove the unincorporated fluorescent ddNTPs. If you are using the AWCGS Capillary Separation service and want to use the Agencourt CleanSeq magnetic bead system for cleanup, you can purchase it direct from Beckman Coulter, visit www.agencourt.com for details of the method. With this method the cleaned products are resuspended in 0.01mM EDTA buffer, and you submit the samples to the AWCGS resuspended in 0.2ml tubes.

NOTE: Beckman Coulter are the suppliers of the Agencourt CleanSeq magnetic bead system in New Zealand.

The '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 will do their sequencing reactions in a 96-well reaction plate, and submit the dried down plate of sequencing reactions to the AWCGS for capillary separation.

Please follow the guidelines in the sequencing kit protocols if you are doing your own sequencing reactions for the 'Plate Service', to get optimal results.

The sequencing kit protocols, which the AWCGS support, can be downloaded as PDF files from the section called 'Chemistry Guide Downloads'. Also refer to the section called 'Tips on Performing Sequencing Reactions'.

The AWCGS 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 AWCGS by e-mail to r.fong@massey.ac.nz if you want to use our 'Plate Service', and we will send you the plates. If you do not fill all 96-wells in the plate with sequencing reactions, you will still be charged the full plate price. Please refer to the section called 'Prices' for details on sequencing using the 'Plate Service'.

Courier of Samples for the ABI Sequencing Service

Prepaid courier bags will be mailed out free of charge to external domestic clients. If you require a bag, you should send an email to Joy Wood at J.R.Wood@massey.ac.nz, stating the number of courier bags to be sent, as well as your courier address, postal address and contact phone number.

Samples are to be couriered to the following address:
Allan Wilson Centre Genome Service
Massey University
Science Tower D, Level 3
Riddett Road
Palmerston North
New Zealand
Attention: Richard Fong