Next Generation Sequencing Instructions

  • Genomic DNA prep: DNA should be of high quality, at least 0.1 µg with a concentration of 10 ng/µl or higher in TE.
  • ChIP DNA prep: at least 1 ng ChIP enriched DNA in 15 µl or less of water, in a low retention tube.  The ChIP DNA should be mostly shorter than 500bp.
  • RNA-Seq Directional Library Prep: at least 0.5 ug total RNA of good quality in 50 µl or less RNase-free water, RNA Integrity Number (RIN) from Bioanalyzer QC should be 9.0 or higher.
  • Small RNA-Seq Library Prep:  at least 0.1 µg total RNA in 10 µl or less RNase-free water, preferably 0.5 -1 µg input total RNA.
  • RNA-Seq Low Input Non-Directional: one or more eukaryotic cells, please contact the Facility Director for recommendations.
  • 10X Genomics Single Cell RNA-Seq V2 library prep:  A single cell suspension of viable cells in compatible buffer, please see the 10X Genomics Single Cell Protocols for recommendations.
  • Pre-made Libraries: A minimum of 20 µl of 5ng/ul pre-made library in EB (Tris-Cl 10mM, pH 8.5) must be provided in a low retention tube. Bioanalyzer QC of library size is highly recommended for each library (before pooling multiplexed libraries).
  • Custom Primer: Custom sequencing primer concentration should be at 100 µM, with a minimum volume of 20 µl.

NOTE:  When filling out the NGS form in iLAB all "Sample IDs" entered should match what you have written on your tubes.  Sample IDs should be short, unique and meaningful.