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Shotgun Sequencing

Shotgun Sequencing  Using the AB 3730xl 

Genome or Large Construct Sequencing

AGRF's current strategies for sequencing large inserts from BACs, fosmids / cosmids, large plasmids and whole genomes utilises a shotgun cloning approach.  Two vectors are currently used; one for general purpose cloning of inserts of up to 5Kb, and the other is for more complex genomes also with an insert range of up to 5Kb.  Larger inserts may be cloned but the limit is 8Kb.

 

The AGRF provides a comprehensive NATA accredited shotgun sequencing service.  Depending on your requirements this service may include:

  • 1x coverage – sequence skimming (minimum)
  • 8x sequence coverage (maximum)

 

The Shotgun Sequencing service is a large scale service with automated pipelines offered for a minimum of 384 samples/clones.  The cost of this service is dependent on the size of the insert or genome, the complexity, accuracy and level of redundancy required for the sequenced material.  There is a Bioinformatics component involved in all levels of the Shotgun Sequencing service.  Our Bioinformatics team can provide various analysis & results files (see Results Formats) made available in electronic format via our secure website.

Shotgun Sequencing:        Sample Submission Requirements

                                            Quote Request
 

Shotgun Sequencing Service Process

Shotgun Seq Flow

 

Shotgun Sequencing using the Roche GS FLX

Shotgun libraries for the GS-FLX are prepared from bacterial/yeast, BAC, cDNA and a range of DNA material with an average insert size of 500bps.  In addition to the whole genome shotgun libraries, there is also a paired end library application.  This application is ideal for aligning contigs generated from the whole genome shotgun approach enabling genome closure to proceed more rapidly.

 

Shotgun Sequencing Service Process

 


 

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