Submission of gDNA for Library Preparation

The Mini-Core will accept high quality gDNA extracts for library preparation and sequencing.

• Input DNA quantity and volume should meet or exceed the following requirements. Submitting sample amounts higher than the above requirements will improve the success of generating high quality libraries.

  • We cannot accept degraded DNA samples. DNA should be of high molecular weight (>15kb) and show no significant fragmentation (e.g. no DNA smear on an agarose gel). See below for characterization requirements.

  • For species with a genome size smaller than 0.5Gb:

    • Quantity: > 125ng

    • Volume: > 25µl

    • Concentration: > 5ng/µl

  • For species with a genome size greater than 0.5Gb:

    • Quantity: > 500ng

    • Volume: > 25µl

    • Concentration: > 20ng/µl

• EDTA concentration should be less than 1mM

  • An EDTA cleanup of extracted DNA can be performed at an additional cost, if necessary.

  • Please indicate the elution buffer used if you are unsure of the EDTA concentration.

• All submitted gDNA samples should be accompanied by two quality checks done by the submitting lab. An estimate of both quantity and quality for HMW gDNA for each sample must be submitted prior to samples being shipped to the CCGP Mini-Core, or additional QC costs will be incurred.

1. Concentration:

   • A fluorometric quantitation (e.g.. Qubit, PicoGreen)

2. Evidence of HMW gDNA:

   • A visible, clear, and labelled band should be present for each sample submitted via a gel image, OR

   • Results from a Bioanalyzer should be submitted for each sample, demonstrating a clear peak for HMW gDNA with clearly labeled sample names