SGT Submission Requirements

pipette into a plate

Our submission requirements have been carefully optimized for experimental success and efficiency. Researchers may only submit samples once an order is placed.  

To minimize workflow disruptions for everyone’s projects, we will return submitted samples that do not adhere to the following requirements:

  • Suspend samples in dH2O, 10mM Tris, or Qiagen EB. Samples in high salt buffers will be cleaned for an extra charge as salt interferes with library preparation.
  • Load samples on a clear semi-skirted polypropylene PCR plate going down the columns (Sample 1 = A1, sample 2 = B1, sample 3 = C1, etc.) as depicted above.
    We do not accept full-skirted, deep well, round-bottom, or flat-bottom plates.
  • Label plates with the order number and sample number range (e.g 4596_S01-S96) and seal firmly with a foil plate seal capable of withstanding -80o C temperatures. If you have more than one plate, number the plates (e.g. 4596_S01-S96_Plate1, 4596_S97-S192_plate2). 
  • If you submit one sample, you may submit in a 1.5mL Eppendorf tube with your order number written on the top of the tube.

In addition to the plating requirements above:

Library Preparation Type Library Prep Kit Used Volume Range (µl) Input Range (ng)
DNA-Seq with Shearing Kapa Hyper Prep 26 - 56 200 - 1000
DNA-seq (full volume reaction) llumina DNA Tagmentation -96 41 - 71 1 - 500
DNA-Seq (1/8 volume reaction) llumina DNA Tagmentation -384 samples 14 - 26 12 - 500
IDT Human Exome (12 plex) Kapa Hyper Prep + IDT Panel 26 - 56 200 - 1000
Twist Human Methylome Twist Methylome  50 225
Methyl-Seq TruSeq DNA Methylation Prep 26 - 58 200 - 1000
ChIP-Seq Kapa Hyper Prep 16 - 56 200 - 1000
Mate-Pair Nextera Mate-Pair Sample Prep 26 - 54 4000+
Custom panels and targeted sequencing contact sequencing@duke.edu 26 - 47 200 - 5000
Oxford Nanopore contact sequencing@duke.edu 26 - 57 1000, 80% > 40 kb
Single cell Tapesteri - Targeted DNA-seq library prep contact sequencing@duke.edu    

In addition to the plating requirements above:

Library Preparation Type Library Prep Kit Used Volume Range (µl) Input Range (ng) RIN
polyA tail enrichment - stranded mRNA-seq Kapa Stranded mRNA-seq kit 26 - 56 200 - 1000 ≥ 7
Ribosome depletion stranded RNA-seq Illumina Tru-Seq Stranded Total RNA-seq kit W/ Ribo-Zero 16 - 20 200 - 1000 ≥ 2
Ribosome and Globin depletion stranded RNA-seq Tecan Universal Plus mRNA-seq 16  - 56 50 - 1000 ≥ 7
Low RNA input RNA-Seq - Not stranded Clontech SMARTer Ultra-low input mRNA-seq kit followd by Kapa Hyper Prep 16 - 20 1 to 10 ≥ 7
small and microRNA-Seq QiaSeq miRNA Library Prep 16 - 56 1 to 10 ≥ 7
FFPE and degraded RNA contact sequencing@duke.edu  

We accept prepared libraries for sequencing; however, we do not guarantee the sequencing results for any customer prepared libraries. 

In addition to the plating requirements above:

Sequencer Flow Cell Min Concentration (nM) Min Volume (ul)
MiSeq -- 10 20
NextSeq -- 10 20
NovaSeq S-Prime 20 30
  S1 20 30
  S2 20 45
  S4 20 100
  Single Lane 10 15

Blood Samples for RNA extractions (2.5ml) should be collected in PAXgene Blood RNA Tubes. Immediately after blood collection, gently invert the PAXgene Blood RNA Tubes 8–10 times.  Store the PAXgene Blood RNA Tubes upright at room temperature (18−25 ̊C) for 2 - 72 hours before transferring to −20 ̊C.  If tubes are to be kept at temperatures below −20 ̊C, freeze them first at −20 ̊C for 24 hours, then transfer them to −80 ̊C.  Tubes should be delivered to us frozen.  Note that the frozen PAXgene Blood RNA Tubes are very fragile and break easily. Tubes must be clearly labeled with your DUGSIM order number and sample number (“4596_S001”). For human blood samples, please make sure to provide us with your IRB number. 

Cells for RNA extractions should be pelleted and snap frozen in Eppendorf or Sarstedt 2.0 mL tubes with liquid removed. Cell pellets should have between 1-3 million cells.