Shotgun Libraries

• Libraries created from precious samples.
• No deletions or missing clones.

• BAC/Fosmid Libraries
  • Random Shear BAC
  • Novel (large insert) Fosmid
  • Partial Digestion BAC
• Sequencing
  • NextGen Sequencing
  • BAC end Sequencing
  • BAC Shotgun Sequencing
  • Sanger DNA Sequencing
• Other Libraries/Cloning
  • Difficult Genome
  • Large/Difficult Insert Cloning
  • NanoClone® & GapFree Shotgun
  • Metagenomic
• Library Screening & Tools
  • Pooling & Superpooling
  • Gridding & Colony Filters
  • Colony Preparation
  • Library Screening
• DNA Prep
  • BAC Miniprep / DNA Prep
  • HMW Genomic DNA Prep

Many laboratories worldwide depend on Lucigen's NanoClone^® and GapFree™ services for economical construction of high accuracy, unbiased shotgun libraries. In fact, Lucigen has become the leading provider of custom shotgun libraries, particularly with “difficult” DNA. Lucigen scientists have discovered numerous novel genes from phage and viral metagenomic DNA isolated directly from boiling hot springs¹.

NanoClone Shotgun Libraries
Access entirely new genomes with Lucigen's NanoClone technology! With as little as 1 nanogram of target DNA, Lucigen will generate a library of >1 x 10⁵ plasmid clones. Trace amounts of gel-separated chromosomes, genomic DNA, uncultured environmental microbes, or base-modified phage are sufficient to generate large plasmid libraries (see Table 1 below). Many of these samples yield few or no clones using conventional techniques, which often require 5-10 µg of starting material.

The NanoClone Library Construction Service includes:

• Size-specific, random physical shearing (HydroShear™ fragmentation)
• High efficiency cloning into a pSMART^® or other Lucigen vector
• QC analysis to confirm size and sequence identity of randomly sampled clones, as well as overall cloning efficiency

Table 1. Examples of genomic libraries prepared using the NanoClone technology

GapFree Shotgun Libraries
GapFree cloning uses Lucigen's revolutionary pSMART transcription-free vectors (US Pat. 6,709,861) to achieve high cloning efficiencies and unprecedented insert stability. As a result, cloning gaps, sequencing gaps, and deleted or rearranged sequences are dramatically reduced. We have developed innovative solutions for cloning a variety of previously "unclonable" DNAs, including:

• Toxic coding sequences
• Strong promoters
• AT- or GC-rich DNA
• Modified bases
• Repeats or hairpins
• Large fragments (>10 kb)
• Trace amounts of precious sample

Lucigen is the exclusive supplier of custom shotgun libraries prepared with the CloneSmart^® technology.

Figure 1. Construction of GapFree DNA Librairies

What you get with the GapFree Shotgun Library Construction Service
As shown in Figure 1, Lucigen will shear, end repair, size select, ligate, and transform your DNA sample to construct a small or large insert, unbiased and highly complex genomic library. We guarantee >50,000 recombinants per library, but a typical library yields >500,000 recombinants.

¹ Schoenfeld T, Patterson M, Richardson P, Wommac E, Young M, Mead DA. (2008) Assembly of Viral Metagenomes from Yellowstone Hot Springs. Appl Environ Microbiol. 74:4164-74.