Sample Submission Requirements
Sample Submission requirements for AGRF services:
Sequencing
AGRFs routine sequencing service encompasses capillary separation (electrophoretic separation only), Purified DNA and Unpurified PCR Product (PD+).
Relevant documents for these services include:
Capillary Separation
The Capillary Separation service is provided for those clients who have completed the DNA labelling (sequencing) reaction and subsequent cleanup themselves and send the dried down pellet to be analysed on the AB3730xl sequencing platforms. Click here for the Capillary Separation Service Workflow.
Templates accepted: ds DNA, PCR product
Chemistry accepted: ABI BigDye Terminator version 3.1
ABI Prism BigDye Terminator Sequencing Reaction Kits
BDT order form and Purchase orders can be faxed to us on (07) 3365 1823.
Purified DNA
The Purified DNA service is provided for those clients who request that AGRF performs the DNA labelling (sequencing) reaction and cleanup. AGRF is supplied with purified template and primers, DNA labelling (sequencing) reaction and cleanup occurs followed by analysis performed on the AB3730xl sequencing platforms. Sequence data delivery is via the AGRF's secure FTP site. Click here for the Purified DNA Service Workflow.
Templates accepted: Plasmid DNA or PCR Product
Chemistry accepted: ABI BigDye Terminator Version 3.1
Unpurified PCR Product (PD+)
The Unpurified PCR Product (PD+) service is designed for users requiring high-volume purification and sequencing of PCR amplicons. AGRF is supplied with unpurified amplicon and primers. Amplicon purification, DNA labelling (sequencing) reaction and cleanup occurs followed by analysis performed on the AB3730xl sequencing platforms. Sequence data delivery is via the AGRF's secure FTP site. Click here for the PD+ Service Workflow.
Samples accepted: PCR Product
Chemistry accepted: ABI BigDye Terminator Version 3.1
PCR re-sequencing
Click here for the PCR Re-Sequencing Service Workflow
DNA source accepted: Genomic DNA (gDNA)
Templates accepted: Purified genomic DNA or glycerol stock / agar stab / bacterial pellet of Bacterial sample/s
Sequencing options available: ABI BigDye Terminator reactions with amplimer sizes between 300 and 600 bp in length
Shotgun Sequencing
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Large insert and Genome sequencing AGRFs current strategies for sequencing large inserts from BACs, cosmids, P1s, PACs, plasmids and from chromosomes and genomes utilises either a shotgun or transposon insertion cloning/sequencing approach. This service may include DNA preparation from clones, shearing/cloning of DNA (shotgun library construction), random sequencing reactions, primer walking and finishing reactions as required as well as contig assembly. Cost is dependent on the size of the insert or genome, the complexity, accuracy and the level of redundancy required for the sequence. An estimate of the percent of clone sequenced for different levels of coverage can be obtained from the Lander-Waterman calculations.
DNA source accepted: BACs, PACs, P1, cosmids, plasmids, chromosomes, genomic DNA |
