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BAC Library Screening

For the purpose of gene targeting vector construction, mouse BAC genomic libraries have advantages over other types of genomic libraries. Because the average size of insert for BAC clones is around 150 kb, the chance is high for having all the necessary pieces in one BAC clone. Another advantage is that many mouse genomic BAC libraries are commercially available.

The purpose for BAC library screening is to identify the clone ID of the clone containing the gene of interest so that we can order it for targeting vector construction.

Because a unique and pure southern probe is critical for the success of BAC library screening, a rigorous procedure is required for making the probe.

A typical BAC library screening involves the following steps,

  • Design the probe primers within the gene of interest (GOI)
  • PCR the probe using the matching genomic DNA as the template
  • Clone the PCR fragment into a vector
  • Sequence the vector insert to confirm that its sequence matches that of the GOI
  • Label the probe with P32
  • Southern blot of the library filters
  • Interpret the result

    To find out more details, please send us a service inquiry/request

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