PacBio Sequencing

The UO GC3F now offers Pacific Biosciences (PacBio) sequencing services on our newly installed PacBio Sequel system!  Single Molecules of native DNA are sequenced in Real Time on the Sequel instrument utilizing devices called "SMRT cells".  Each SMRT cell can yield between 5 and 10 Gb of sequence data, with a read length of greater than 10 kb on average.  

Curious about how this works?  Watch this.

 

Pricing:

Sample Preparation *  
Internal Rate (per library) $450.00
External Rate (per library) $675.00
Sequencing  
Internal Rate (per SMRT cell) $800.00
External Rate (per SMRT cell) $1,200.00

*Obtaining sufficient quantities of high molecular weight DNA for PacBio sample preparation can be difficult.  Additional service fees (Fragment Analyzer, Pippin Prep, etc.) may apply for extra QC and troubleshooting.

____________________________________________________________________________________________

IMPORTANT NOTE FOR USER-PREPPED LIBRARIES:

If you are planning to prep your own PacBio libraries, please be sure to ship them frozen on dry ice. Jostling of thawed samples during shipping may damage your libraries and prevent sequencing. 

____________________________________________________________________________________________

IMPORTANT NOTES FOR GC3F-PREPPED LIBRARIES:

If you are planning to provide the GC3F with DNA for us to make the PacBio libraries for you, the quality of the input DNA is extremely critical for ensuring adequate library yield and high sequencing quality. Please be sure your purified dsDNA meets the following strict PacBio requirements:

  • All steps of DNA extraction, purification, and storage performed in Eppendorf LoBind tubes
  • Submit a minimum of 5 µg dsDNA (ssDNA cannot be prepped)
    • Must be quantified using a fluorometric assay such as Qubit or Quant-iT (spectrophotometric assays are not adequate)
    • Be sure to very thoroughly (but very gently!) mix your sample using wide-bore tips prior to taking an aliquot for quantification
    • If you have less than 5 µg, please contact us to discuss options
  • DNA is of high quality = 
    • Fragments are of sufficient length for your desired sequencing application (contact us for details on your specific project)
    • No RNA present
    • Has not been exposed to intercalating dyes, UV radiation, ethidium bromide, high temperatures, extreme pHs, or vortexing
    • Minimal nicks, breaks, and other damage
    • Minimal time spent stored at 4°C
    • Minimal freeze-thaw cycles
    • If shipping to our facility, must be sent frozen on dry ice
  • DNA is highly purified =
    • Purified and stored in EB or TE buffer
    • 260/280 nanodrop ratio = at least 1.8
    • 260/230 nanodrop ratio = between 2.0 and 2.2
    • No visible particulates, precipitates, colors, cloudiness, or viscous substances

____________________________________________________________________________________________

Please contact Doug Turnbull or Maggie Weitzman for further details on PacBio sample submission. Thank you!