Construction of mouse 129/Ola BAC library for targeting experiments using E14 embryonic stem cells

Area of Science:

• Genomics
• Molecular Biology
• Biotechnology

Background:

• Gene targeting enables specific gene modification.
• 129 mouse strain embryonic stem (ES) cells are ideal for gene targeting due to ease of culture.
• ES cell-derived genomic clones enhance homologous recombination frequency in gene targeting.

Purpose of the Study:

• To generate an arrayed mouse 129/Ola bacterial artificial chromosome (BAC) library using E14.1 ES cells.
• To provide a valuable resource for gene targeting and genome analysis.

Main Methods:

• Generation of an arrayed mouse 129/Ola BAC library from E14.1 ES cells.
• Isolation of over 135,000 BAC clones with an average insert size of 110 kb.
• PCR screening of BAC clones for efficient locus identification.

Main Results:

• The library comprises >135,000 BAC clones with a mean insert size of 110 kb.
• Estimated 5.5-fold coverage of the mouse genome.
• BAC clones are screenable via PCR within 2 days.
• All 8 examined loci are present in the library.

Conclusions:

• The developed BAC library is a valuable resource for gene targeting studies utilizing E14 ES cells.
• The library supports efficient genome analysis and facilitates precise gene modification strategies.
• This resource aids researchers in advancing gene targeting experiments and understanding genomic structures.