Sample Submission Requirements
Sample Submission requirements for AGRF services:
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
How to Prepare Your Capillary Separation (CS) Samples (pdf)
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
How to Prepare Your Purified DNA (PD) Samples (pdf)
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
Plasmid / Clone Sequencing
AGRF routinely sequences cDNA/EST libraries provided by customers. This service includes:
- plasmid DNA template preparation
- DNA labelling (sequencing) reaction
- cleanup
- gel separation run
- sequence data delivery by FTP
DNA source accepted: cDNA/EST libraries constructed in pUC-derived and other high copy plasmid vectors.
Templates accepted: Glycerol stocks, agar stabs or bacterial colonies
on agar plates to be processed in batches of 96 to allow for use of
high throughput DNA preparation protocols and robotics.
Sequencing options available: ABI BigDye Terminator with either standard M13 standard sequencing primers or custom primers.
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 |
