Sample Preparation Methods for ABI 3130xl

PCR Purification Methods:
  • ExoSap (recommended)
    Exo-Sap Purification will eliminate residual primers and dNTPs. This procedure will NOT eliminate unspecific bands or primer dimers.
    • Prepare a Master Mix of the following reagents:
      0.30 uL of 20U/uL Exonuclease I
      0.45 uL of 1U/uL Shrimp Alkaline Phosphatase
      2.25 uL ddH2O
    • Add 3 uL of Master Mix to <6-10 uL PCR product.
    • Cycling: 37˚C – 35 mins, 80˚C – 15 mins, 4˚C – 1 min.
    • Store at -20˚C until until ready to bring to the facility.
Cycle Sequencing & Purification Protocol:
We have our own custom cycle sequencing protocol. If you are interested in protocol details for Ready to Load submission, please contact us.

SGF Protocols for NGS
Below you will find a list of protocols developed at SGF from different sources. The attachments are at the bottom of the page.
  • Single-Digest RAD Tag Sequencing Library Preparation
    Our protocol is based on Etter et al. 2011 MMB 772:157-78 and on Baired et al. PloS ONE 3:e3376; with suggestions and quantities from Ximena Vélez from Papa Lab UPR-RP. It incorporates the use of the BluePippin (Sage Sciences) for Size Selection, as optimized by Silvia Planas, SGF Staff, AMPure purification procedures, and BioAnalyzer traces for Quality Control. It was written by Dania Rodriguez, SGF Staff.

Jun 10, 2015, 8:09 AM