Home » Services » Sequencing » Sample Submission Requirements

Sample Submission Requirements

Sample Submission requirements for AGRF services:

  • Capillary Separation

  • Purified DNA

  • Unpurified PCR Product

  • PCR re-sequencing

  • Shotgun Sequencing


  • Sequencing


    AGRFs routine sequencing service encompasses capillary separation (electrophoretic separation only), Purified DNA and Unpurified PCR Product (PD+).

    Relevant documents for these services include:

  • Guide to AGRF Routine Sequencing Service (PDF)

  • BDT Order Form (PDF)


  • 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

    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

    Templates accepted: Purified genomic or clone DNA, glycerol stock/ agar stab (BACs only accepted as glycel/agar stabs)

    Sequencing options available: Random sequence coverage of DNA of interest ranging from 1x (sequence skimming) to 10x to obtain full-length double-stranded sequence.

     

    Back to top 

    Recent News