Whole Genome Shotgun Libraries

A Whole Genome Shotgun library is an integral tool in efficiently generating DNA sequence (Sanger) from total genomic DNA.

Amplicon Express (AEX) uses an optimized Hydroshear technology (random mechanical shearing and end repair) to generate insert DNA minimizing cloning bias. AEX Whole Genome randomly sheared shotgun libraries are cloned in a high copy plasmid vector that's easily sequenced with common primers and proven stable.

AEX can construct libraries with very specific insert sizes, determined by the client's scientific goals (i.e. 1Kb +/- 250 bp or 2Kb, 3Kb, 4Kb, 6Kb, 8Kb, 10Kb). AEX will provide living, frozen clones arrayed on 96 or 384 well plates, ready to prep and sequence. AEX shotgun subclones can be end sequenced and/or can be primer walked to close gaps in a cost effective manner. Having shotgun clones gives a distinct advantage over Next Generation sequencing, because there is a resource of cloned fragments for gap closure (after the initial assembly is completed).

If desired, AEX can prepare DNA, sequence and assemble the clones into contigs across the genome.

When to choose a Whole Genome Shotgun Library:
  1. When Sanger sequencing is the technology of choice for a DNA sequencing project.
  2. In conjunction with 454 or other Next Generation technologies 1x to 3X coverage in Sanger reads on Whole Genome Shotgun library clones gives a resource for gap closure (primer walking clones containing gaps).
  3. After a genome has been sequenced using 454 or other Next Generation technologies and gaps still exist across important regions. Shotgun clones can be arrayed onto nylon filters, screened with probes from gap frontiers and primer walked to close gaps.


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