Library Submission Guidelines

New York
Abu Dhabi

How to Prepare Libraries for Submission:

Labs are responsible for quantifying their own libraries using qPCR (reagents are purchasable from the GenCore) and pooling libraries prior to submission. Please follow these steps:

General Instructions:

  • Quantify stock libraries with qPCR and adjust the concentration of library (according to the table below) using Tris-Cl 10mM, pH 8.5 with 0.1% Tween 20. Pools must be submitted in Eppendorf LoBind low retention tubes.
  • If you intend to run a single library on a lane: submit 25 uL of  library in the concentration specified below. This will constitute the “library pool” for that lane.
  • If you intend to run two or more libraries in a lane: combine equal/unequal volumes of diluted libraries (according to the table below) and submit 25 uL of this pooled library mixture to the core for sequencing. (Note: pooling can be completed at equal or unequal distributions depending on the requirements of a given experiment).
  • To avoid concentration changes associated with freeze-thaw, libraries should not be frozen before dilution, pooling or submission to GenCore.
  • Libraries must be submitted no more than 5 days prior to the start of the sequencing run. We will store libraries at 4°C to prevent concentration changes associated with freeze-thaw, so we don’t want to receive the samples too early. Please keep excess library in your lab freezers.
Illumina HiSeq 2500 Illumina NextSeq 500 Illumina MiSeq
2 nM 4 nM 4 nM

*Note: If the library pool concentration is below the submission concentrations listed in the above table, contact GenCore for further instructions and advice.

Nextera & Nextera-like Libraries:

  • Libraries prepared with Nextera or Nextera-like protocols (ie ATAC-seq) must be denoted at time of submission by checking the Nextera box in TuboWeb or indicating “yes” for Nextera in the csv import under the Pool Information.
  • If submitting libraries for a HiSeq 2500 run, be sure to contact the GenCore in advance.  This library type requires additional primers and reagents. GenCore needs ample notice to make sure to have these in stock in time for the run.
  • When using the Nextera XT kit, store samples at -20 degrees before bead normalization. Once this is completed the sample is only stable for ~1 week. It is best to wait until the run is scheduled and the run confirmation e-mail has been sent by the GenCore to perform this last step of the protocol.

Custom Sequencing Primers:

  • If using custom sequencing primers, these must be submitted along with samples.
  • Aliquots must be at 100 uM in at least 20 ul
  • Primers must be diluted in EB or DI Water
  • Must be in a Low-bind tube and clearly labeled