MiSEQ

PacBio

Get the Best of Both NGS Worlds:  “PACBIO Read Length” and “ILLUMINA Read Depth”.

Many researchers want detailed and accurate sequence of just a few BAC clones, from a few regions, not the entire genome. We have the ideal strategy to make that happen.

Inspired from our clients’ needs and requests, our method utilizes the strength of both PacBio and Illumina sequencing platforms for individual BAC clones. The data yield is typically between 50X and 150X coverage for each BAC clone comprised of both PacBio 20Kb library reads and MiSeq 500 to 700bp Paired-End reads.

We are able to accomplish this inexpensively, $800 per clone, because we make a single PacBio library by combining 12 BAC clones provided by one or more clients. We sequence that library on a PacBio machine and then assemble the data. In order to identify individual clones, we also make 12 individually tagged Illumina libraries, which are sequenced on a MiSeq instrument. We then assemble & map the Illumina data with the PacBio data connecting the BAC clone name to each contig, scaffold or assembly.

Strategic Overview:

  1. Make High Quality NGS grade BAC DNA (low E. coli genomic DNA).
  2. Combine 12 pooled BAC Clones in one NGS PacBio 20 Kb library.
  3. Prepare individual Paired-End libraries for each BAC clone and run on a MiSeq instrument.

We are getting excellent results with this strategy and would like to do it for your BAC clones!

Contact us to get a BAC Clone Sequencing quote.