Random Shotgun/Subclone Libraries

Shotgun libraries are an effective and economical tool for efficiently generating DNA sequence (Sanger) or identifying and studying small specific regions of interest (genes, promoters or other factors) from cloned DNA contained in a BAC, Fosmid or Cosmid vector.

Amplicon Express (AEX) uses an optimized Hydroshear technology (random mechanical shearing and end repair) to generate insert DNA minimizing cloning bias. AEX 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. 2Kb +/- 250 bp or 4Kb, 6Kb, 8Kb, 10Kb). AEX will provide living, frozen subclones 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 with insert sizes of the appropriate range can reduce the difficulties of repeats found in a large (+100 Kb) genomic clone.

If desired, AEX can prepare DNA, sequence and assemble the subclones into contigs covering the original clone insert.

When to choose a Random Shotgun Library:
  1. When Sanger sequencing is the technology of choice for a DNA sequencing project.
  2. After BAC's have been sequenced using 454 or other Next Generation technologies and gaps still exist across important regions. Shotgun subclones can be primer walked for gap closure.
  3. When a BAC insert or other clone has many repeat elements that are difficult/impossible to resolve using Next Gen sequencing technologies (especially Illumina and ABISOLiD with short reads).


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